ImagineNano Abstract Booklet (part II) by Phantoms Foundation

Oral Development of Hydrogel membranes for long term biopotential monitoringusingflexible wet electrodes

Carme de Haro1 2 R.Ruiz 1 P. Lacharmoise and 1 N. Guilera

cdeharo@cetemmsa.com 1 2

Fundació CETEMMSA, Avd. Ernest Lluch, 36 08302 Mataró (Barcelona) RGB Medical Devices S.A.,Alfonso Gómez 42 28037 Madrid - Spain

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Ubiquitous monitoring of biopotentials can greatly enhance comfort, health and safety of patients dealing with chronic diseases. For home use, clinical grade adhesive electrodes are often cited as irritating and uncomfortable, leading to low usage compliance [1]. For long-term monitoring it is mandatory to have innocuous and disposable electrodes with constant impedance over long periods of time. In previous work we developed 100% dry electrodes to fabricate a T-shirt for long-term ECG monitoring [2]. These dry electrodes exhibit low stability: they change their impedance over length of use and may even vary within a single ECG register. We concluded that dry electrodes can´t bring enough stability and thus electrodes with interfacial wet membranes are preferred. In the current work we present the development, fabrication and characterization of wet bioelectrodes for long-term biopotential monitoring. The new printed and textile-based electrodes allow carrying out long-term recordings due to their biocompatibility (non-toxic, nonallergic and non-irritant). However, they present high electrode-tissue interface impedance, providing a low level signal with clear motion artifacts that can´t be digitally filtered without losing valuable information. To improve the signal quality, a non-irritating hydrogel membrane which improves the contact impedance and reduces the movement artifact is being developed. Apart from biocompatibility, the developed membranes must satisfy two basic requirements: good mechanical adhesion to the skin of the patient and interfacial impedance equal or lower than conventional ECG electrodes. We are studying two different hydrogel formulations based on acrylamide polymers on one hand and polyvinylpyrrolidone (NVP) on the

other hand. Apart from the hydrogel formulation, we focused our attention in the mechanical behavior of the membranes. In order to improve their mechanical stability we propose the use of embedded tissue that strengthens the hydrogel membrane without losing flexibility. A wide variety of tissues were tested, being XXX and XXX the ones that showed best compatibility into the membrane. Pictures of tissue embedded membranes are shown in Figure 1. Relative impedance measurements between commercial ECG electrodes and textile electrodes with different formulations of interfacial membranes are shown in Figure 2. The impedance of the textile electrodes remain below the measured impedance for commercial ECG electrodes independently of the NVP formulation membrane. We have proved that hydrogel-based interfacial membranes can be used in combination with textile electrodes for long term ECG monitoring. The developed solution provides good functional behavior in terms of stability, mechanical adhesion and conductivity. These membranes are actually being tested in T-shirt prototypes with integrated textile sensors which are expected to allow for comfortable ubiquitous ECG monitoring. The final product is a T-shirt where the electrodes will do the register in continuous. The T-shirt has 4 different register points, two on the front of the Tshirt and the others two on the back. In Figure 3 we present one prototype of the T-shirt with textile ECG electrodes.

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References [1] A. Gruetzmann, S. Hansen, J. M端ller. Novel dry electrodes for ECG monitoring. Physiol. Meas. 28 (2007) 1375-1390. [2] C. de Haro, C. Carenas, R.Ruiz, E. Calderon, L. Francesch. Development of new printed electrodes for biopotential monitoring. Tarragona 25-27 May 2012, 8th International Conference on Organic Electronics -ICOE 2012. Oral. [3] L. Beckmann, C. Neuhaus et al. Characterization of textile electrodes and conductors using standardized measurement setups. Physiol. Meas. 31 (2010) 233-247.

Figure 1: a) hydrogel membrane with synthetic wadding. b) hydrogel membrane with web cotton c) hydrogel membrane with silk.

Figure 2: Impedance register of different ECG electrodes, referenced to ECG commercials.

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Figure 3: View of different parts of developed T-shirt to biopotencials registers.

Invited Force nanoscopy of microbial pathogens

Yves DufrĂŞne

Yves.Dufrene@uclouvain.be Institute of Life Sciences, UniversitĂŠ catholique de Louvain, Belgium

Progress in nanomedicine relies on the development of advanced tools for imaging and manipulating biological systems on the nanoscale. Among these, atomic force microscopy (AFM) techniques have emerged as a powerful platform for analyzing the structure, properties and functions of living cells, including microbial pathogens. AFM imaging enables researchers to observe microbial cell walls in solution and at high resolution, and to monitor their remodelling upon interaction with drugs. In addition, single-molecule force spectroscopy analyzes the localization, mechanics and interactions of the individual cell wall constituents, thereby contributing to our understanding of how cell surface receptors are spatially-organized (e.g. clustering) and respond to force (e.g. single specific bonds, sequential unfolding of single protein domains, zipper-like adhesion, and spring-like properties). Knowledge of these molecular properties is critical to our understanding of pathogen surface interactions. In the future, we expect this new form of nanoscopy to have an important impact on nanomedicine, particularly for understanding microbe-drug and microbe-host interactions and for developing new antimicrobial strategies. I will survey recent breakthroughs we have made in applying AFM to microbial pathogens [1-4].

References

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[1] Beaussart et al. (2012). ACS Nano 12: 1095010964. [2] El-Kirat-Chatel et al. (2012). ACS Nano 12: 10792-10799. [3] Alsteens et al. (2012). ACS Nano, 6: 77037711. [4] Tripathi et al. (2013). ACS Nano, 10.1021/nn400705u.

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Oral Metallophilic Hydrogels: A New Family of Injectable and Self-Healing Materials

Damien Dupin P. Casuso, N. Diaz I. Odroziola, H. - J. Grande G. Cabañero and I. Loinaz

IK4-CIDETEC, Parque Tecnológico de San Sebastián, Paseo Miramón 196, 20009 Donostia-San Sebastián, Spain

ddupin@cidetec.es

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properties for a wide range of application from tissue engineering as injectable scaffolds, synovial liquid replacement due its shear-thickening behavior to material for wound healing.

[1] Burkoth, A. K. et al. BIOMATERIALS 21 (2000) 2395-2404. [2] Klouda, L. et al. EUR. J. PHARM. BIOPHARM. 68 (2008) 34-45. [3] Casuso, P. et al. ORG. BIOMOL. CHEM. 8 (2010) 5455-5458. [4] Odriozola, I. et al. ORG. BIOMOL. CHEM. 9 (2011) 5059-5061. [5] Patent number EP11382365. Figures

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References

Figure 1: Self-healing behavior of the supramolecular hydrogel.

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In-situ forming hydrogel systems have attracted considerable interest as injectable scaffolds for tissue engineering and drug delivery due to their easy applications and minimally invasive injection procedure. [1,2] Our research group has recently developed a new generation of injectable supramolecular hydrogels based on low molecularweight Au (I) and Ag (I) thiolates. [3,4] Here we present a new family of polymeric hydrogel with self-healing properties and adjustable mechanical properties. [5] Supramolecular polymeric hydrogels were obtained by mixing an aqueous solution of salt metals (AgNO3 or HAuCl4) with an aqueous solution of thiolated polymer at physiological pH. The instantaneous gel formation allows its application as injectable scaffold for tissue engineering, e.g. bone or cartilage replacement. Interestingly the supramolecular polymeric hydrogel exhibits self-healing properties (Figure 1) due to the reversible supramolecular interaction of thiolates metal (I). Moreover, the mechanical properties of these supramolecular hydrogels could be tuned on demand by varying the amount of metal ions incorporated into the hydrogel. For example, hydrogels with high amount of added gold exhibit a shear-thickening behavior similar to synovial liquid (Figure 2). Furthermore supramolecular hydrogels with lower gold concentration show more toughness but their self-healing properties are kept in (Figure 3). Also a judicious selection of the metal allows the hydrogel to be an excellent cell proliferation scaffold or to have antibacterial properties when gold and silver are used, respectively. In summary, this new family of polymeric supramolecular hydrogels offers advantageous

Figure 2: Variation of G’ (solid behavior) and G’’ (liquid behavior) with frequency.

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Figure 3: Variation of the storage modulus (G’) with different amount of metallophilic interactions ranging from 20 to 110 mol % based on the initial quantity of thiols.

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Oral Nanoassembly of the Protein Corona on Nanoparticles

Giancarlo Franzese 1 P. Vilaseca 1 O. Vilanova M. Bernabei1 2 K. A. Dawson

Cellular responses to materials in a biological medium reflect greatly the adsorbed biomolecular layer, rather than the material itself. Here, we study by molecular dynamic simulations the competitive protein adsorption on surfaces, i.e. the non-monotonic behavior of the amount of protein adsorbed on a surface in contact with plasma as a function of contact time and plasma concentration. We study the effects of different curvatures and nanoparticles size when the surface chemistry is the same [1]. References [1] P. Vilaseca, K. A. Dawson, and G. Franzese, â&#x20AC;&#x153;Understanding and modulating the competitive surface-adsorption of proteinsâ&#x20AC;?, arXiv: 1202.3796 (2012). Figures

gfranzese@ub.edu

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Departament de Fisica Fonamental, Facultat de Fisica, Universitat de Barcelona, Diagonal 645, 08028, Barcelona, Spain, 2 Center for BioNano Interactions (CBNI), University College of Dublin, Ireland

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Figure 1: Simulation snapshot of a model plasma solution in contact with a nanoparticle (in the center). The model plasma is made of three different proteins (Albumin, Immunoglobulin-gamma, Fibrinogen) adsorbing on a hydrophobic nanoparticle and selfassempling in a protein soft corona.

Invited Challenges in spintronic platforms for biomedical applications 1

INESC-MN–Ins. de Engenharia de Sistemas e Computadores-Microsistemas e Nanotecnologias and IN-Ins. of Nanoscience and Nanotechnology, Rua Alves Redol, 9, 1000-029, Lisbon, Portugal; 2 INL–Int. Iberian Nanotechnology Laboratory, Av. Mestre José Veiga, 4715-31 Braga, Portugal 3 IBB-Ins. for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho Campus de Gualtar, 4700-057 Braga, Portugal 4 IBB- Institute for Biotechnology and Bioengineering, Center for Biological and Chemical Engineering (CEBQ), Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisbon, Portugal 5 INESC-ID–Ins. de Engenharia de Sistemas e Computadores-Inovação e Desenvolvimento, Rua Alves Redol, 9, 1000-029, Lisbon, Portugal 6 Laboratory of Neurosciences, Faculty of Medicine, University of Lisbon, Portugal 7 ICVS, U.Minho, Braga, Portugal

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Integrated spintronic biochip platforms are being developed for portable, point-of-care, diagnostic and cytometric applications [1,2]. Hybrid systems incorporating magnetoresistive sensors are applied to neuroelectronic studies and biomedical imaging, namely magnetoencephalography and magnetocardiography. Lab-on-a-chip MR-based platforms are under development to perform biological studies at the single molecule level. This review introduces and discusses the potential and main characteristics of those MR-based biomedical devices, comparing to the existing technologies, while giving particular examples of targeted applications. Applications to the detection of DNA hybridization events (DNA-chips) [3] and antibodyantigen recognition at immunoassays (immunochips) [4] are discussed. Particular examples for cell free DNA and genomic sequences detection, for pathogen (Salmonella enteritidis, see Fig.1) detection and for flow cytometry (separation and counting) of CD34+ magnetically labeled cells coming from bone marrow or cord blood samples are given. Moreover, lateral immuno-assay configurations where analytes are labeled with magnetic nanoparticles are discussed. For biomedical imaging applications, field sensitivity is being pushed towards 1pT/sqrt(Hz) and below in hybrid devices incorporating flux guides with the magnetoresistive element allowing the direct detection of bio-magnetic fields (from brain and heart). For neuroelectronic applications, sensors are being incorporated in microelectrode arrays (Si and polyimide) to record spontaneous or stimulated neural activity (in vitro and in vivo, see Fig.2).

Paulo P. Freitas

1,2

F.A. Cardoso1, V.C. Martins2 E. Fernandes2,3, S.A.M. Martins4 T. Dias1,4, J.P. Amaral1 S. Cardoso1, J. Germano5 T. Costa5, M.S. Piedade5 J. Gaspar2, J. Noh2 M. Costa2, H. Fonseca2 M. Debs2, R. Ferreira2 A.M. Sebastião6 and V. Pinto7

paulo.freitas@inl.int

References [1] P. P. Freitas, F. A. Cardoso, V. C. Martins, S. A. M. Martins, J. Loureiro, J. Amaral, R. C. Chaves, S.Cardoso, L. P. Fonseca, A. M. Sebastião, M. Pannetier-Lecoeur and C. Fermon “Spintronic platforms for biomedical applications”, Lab Chip, 12, 546-557 (2012). [2] V.C. Martins, J. Germano, F.A. Cardoso, J. Loureiro, S. Cardoso, L. Sousa, M. Piedade, L.P. Fonseca, P.P. Freitas “Challenges and trends in the development of a magnetoresistive biochip portable platform” Journal of Magnetism and Magnetic Materials, 322, 1655-1663 (2010). [3] V.C. Martins, F.A. Cardoso, J. Germano, S. Cardoso, L. Sousa, M. Piedade, P.P. Freitas, L.P. Fonseca “Femtomolar Limit of Detection with a Magnetoresistive Biochip” Biosensors and Bioelectronics, 24, 2690-2695 (2009). [4] Sofia A. M. Martins, Verónica C. Martins, Filipe A. Cardoso, Paulo P. Freitas, Luís P. Fonseca, “Waterborne Pathogen Detection Using a Magnetoresistive Immuno-Chip”, in Molecular Biological Technologies for Ocean Sensing (Sonia Tiquia, ed.), Springer Protocols Handbooks series, Humana Press, 263-288 (2012).

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Figure 2: (a) SEM image of the probe shafts with MR sensors, (b) transfer curve of a used MTJ sensors and its structure. (c) MR needles array with respect to the different hippocampus structures and (d) sensor output after an impulse in the CA1 brain region..

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Figure 1: Phage-based magnetoresitive biochip measurements performed on an electronic reader. (a) Measurement curves for different sensors (Reference sensor – no probe; Positive sensor – specific phage for Salmonella; Negative sensor – phage for Campylobacter) on the same chip for a Salmonella sample analysis. (b) Picture from the biochip over a culture plate with grown Salmonella colonies. (c) SEM picture from a gold surface with immobilize bacteriophage recognizing Salmonella cells.

Oral Application of the enzymatic product mediated stabilization of in situ produced CdS quantum dots: serum paraoxonase, acetylcholine esterase and glutathion reductase detection

Gaizka Garai-Ibabe L. Saa and V. Pavlov

ggarai@cicbiomagune.es

CICbiomaGUNE, Parque Tecnologico de San Sebastian, Paseo Miramón 182, 20009, Donostia, Spain

Detection of analytes in biosensing is carried out by interaction of biological recognition elements (DNA, enzymes, antibodies and so on) with analytes of interest. The event of recognition is transduced and amplified to yield a signal measurable by physical techniques such as UV-Vis and fluorescence spectroscopy, Raman spectroscopy, electrochemistry etc. Semiconductor inorganic fluorescent nanoparticles, known as quantum dots (QDs), have been extensively used in recent years as labels for antibodies, DNA and small molecules in bioanalytical affinity assays and imaging.[1,2,3]The advantages of QDs over traditional organic fluorophores include higher quantum yield, reduced photo-bleaching and higher extinction coefficient. Until now QDs based assays relied on pre-synthesized semiconductor NPs. However, the drawbacks of such assays used to be the high background signals caused by the nonspecific adsorption of decorated QDs on surfaces or poor quenching of donor couples. Thus we believed that the generation of QDs in situ can address these drawbacks of relevant analytical systems by decreasing the background signal. Here we introduce a novel concept in bioanalysis and apply it to the rapid, highly sensitive and inexpensive fluorimetric detection of different enzymes with biomedical interest: serum paraoxonase-1(PON-1)(Figure 1), acetylcholine esterase (AChE) (Figure 2) and glutathione reductase (GR)(Figure 3 3).[4,5,6].

[2] Goldman, E. R.; Clapp, A. R.; Anderson, G. P.; Uyeda, H. T.; Mauro, J. M.; Medintz, I. L.; Mattoussi, H. Anal. Chem., 76 (2004) 684. [3] Green, M. Angew. Chem. Int. Ed., 43 (2004) 4129. [4] Garai-Ibabe G., Möller M., Pavlov V. Anal. Chem., 84 (2012) 8033. [5] Garai-Ibabe G., Saa L., Pavlov V. submitted Analytical Chemistry. [6] Garai-Ibabe G., Saa L., Pavlov V. submitted Analytical Chemistry

Figures

Figure 1 A) Detection of PON1 activity by the enzymatic modulation of growth of fluorescent CdS QDs. B) Emission spectra of CdS QDs formed in the system containing PTA (2mM), Na2S (0.3 mM), Na3PO3S (0.9 mM), Cd(NO3)2 (1.25 mM) and various concentrations of rHuPON1: a) 0 mU mL-1; b) 0.625 mU mL-1; c) 1.25 mU mL-1; d) 2.5 mU mL-1; e) 5 mU mL-1; f) 10 mU mL-1; g) 20 mU mL-1; h) 40 mU mL-1. C) Calibration curve of rHuPON1, measured by the enzymatic modulation of growth of fluorescent CdS QDs, □, and the standard PA method, ■.

References

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[1] Zhang, Y.; So, M.-K.; Loening, A. M.; Yao, H.; Gambhir, S. S.; Rao, J. Angew. Chem. Int. Ed., 45 (2006) 4936.

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Figure 3 A) Detection of GR activity by the enzymatic stabilization of fluorescent CdS B) Emission spectra of CdS QDs formed in diluted human serum (1:1000) containing NADPH (17.5 碌M), Na2S (0.1 mM), Cd(NO3)2 (1.25 mM) and different concentrations of GR: a) 0 pM; b) 5 pM; c) 10 pM; d) 25 pM; e) 50 pM;. C) Calibration curve of GR at 位=550 nm.

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Figure 2 A) Detection of AChE activity by the enzymatic productmediated stabilization of fluorescent CdS QDs B) Fluorescence intensities of the formed CdS QDs in the system containing different concentrations of added standard solution of AChE, ATCh (0.5 mM), Na2S (0.1 mM), Cd(NO3)2 (1.25 mM) and varying volumes of human serum a) 5 nL; b) 6.7 nL; c) 10 nL; d)13.3 nL. C) Dependence of the calculated AChE activity on serum volume.

Oral Detection of Alzheimer Disease Markers by High Performance Plasmonic Silver Nanostars

Jose V. Garcia-Ramos A. Garcia-Leis and S. Sanchez-Cortes

jvicente.g.ramos@csic.es Instituto de Estructura de la Materia. IEM-CSIC. Serrano, 121. 28006 Madrid. Spain

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The manufacture of nanoparticles (NPs) with metallic plasmonic properties, displaying advanced spectroscopic applications is one of the tasks of our research Group. In fact, shape control of metal nanostructures is a promising strategy to tailor their physical and chemical properties. A variety of metal nanostructures with well-defined morphology have been obtained e.g. spheres, triangle, cubes, octahedrons, rods, and plates. Recently, metal nanostructures with complex three dimensional (3D) surface morphology, which are often referred to as nanoflowers or nanostars in literature, have received considerable attention due to their excellent performance as SERS (Surface Enhanced Raman Scattering) substrates. [1] Further, for a metal nanostar, the hybridization of plasmons of the core and tips could dramatically increase the excitation cross section and the electromagnetic field enhancements of the tip plasmons, which results in better SERS performance. [2] The main purpose of our work is the fabrication of nanoscopic systems leading to high electromagnetic field intensifications. The strategy for obtaining these systems has been the preparation of anisotropic NPs with special morphologies leading to high field enhancement, as triangular nanoprisms and nanostars. Progress has been made in a very special way to develop protocols for the preparation of silver nanoshaped nanoparticles, which were not prepared so far in silver. These nanoparticles have a larger number of advantages in relation to gold, since Ag is more efficient in SERS, because of the greater intensification factor, and because of their wider range of activity, as the Ag NPs exhibit plasmon

resonances in the visible spectrum and the nearinfrared. In addition, the protocols of preparation of Ag nanostars were aimed at avoiding the use of surfactants, which are normally used in the preparation of Au nanostars, since these substances prevent the approach of the analyte to the surface, thus decreasing the efficiency of these systems in spectroscopy. In this work we describe the one-step colloid synthesis of silver nanostars and their characterization by spectroscopic and microscopic techniques (Extinction spectroscopy, SERS, TEM and SEF). Two nanostar types have been fabricated. One of them by synthesis in one step of chemical reduction of nitrate silver solution with neutral hydroxylamine (NS-Ag). The second one, by synthesis in two steps adding gold solution to the first Ag nanostars and reducing with citrate at 1% at 100ยบC (NS-Ag@Au). Figure 1 shows the TEM images (inset) and the extinction spectra of both colloids. In particular, the NS-Ag@Au nanoparticles have a thin uniform shell of gold coating the silver core. Extinction spectra of these nanoparticles show a big background at high wavelengths indicating a high size and shape distribution with many different numbers of tips, in both cases. The extinction peak about 535 nm in the case of NS-Ag@Au is indicative the presence of gold shell in the nanoparticles. In order to test these nanostar morphologies as plasmon surfaces, Congo Red (CR) and Thioflavine (ThT) dyes were characterized by vibrational spectroscopy (IR, Raman, SERS) and SEF. CR and ThT are used as histological dyes for staining the peptide _-amyloid, which is a pathologically aggregated form of protein and found in some

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Figure 1 Extinction spectra and TEM (inset) images of NS-Ag and NSAg@Au nanoparticles.

Acknowledgements: This work has been supported by the Spanish Ministerio de Econom铆a y Competitividad (grant FIS2010- 15405) and Comunidad de Madrid through the MICROSERES II network (grant S2009/TIC-1476). A.G. - L. also acknowledges CSIC and FSE for a JAE-predoctoral scholarship.

References [1] X.S. Shen, G.Z. Wang, X. Hong, W. Zhu. Phys. Chem. Chem. Phys., 11 (2009) 7450-7454. [2] V. Giannini, R. Rodriguez-Oliveros, J.A. Sanchez-Gil. Plasmonics, 5 (2010) 99-104. [3] T. Miura, C. Yamamiya, M. Sasaki, K. Suzuki, H. Takeuchi. J. Raman Spectrosc., 33 (2002) 530 535. [4] S.A. Hudson, H. Ecroyd, T.W. Kee, J.A. Carver. FEBS Journal, 276 (2009) 5960-5972.

Figure 2 SERS spectra with NS-Ag and NS-Ag@Au without aggregation for (a) RC and (b) ThT. 位exc. = 532 nm. Spectra normalized to the water band.

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human diseases including Alzheimer and prion diseases. [3,4] Figure 2 shows the SERS spectra of RC and ThT obtained using NS-Ag and NS-Ag@Au as SERS substrates. As can be seen, intense SERS bands of both RC and ThT were obtained in the fabricated nanostars. However, the intensity was higher when using NS-Ag@Au. In addition, the SERS enhancement is much higher in the case of RC. The higher activity of the latter nanoparticles is attributed to the higher extinction of light occurring on NS-Ag@Au due to the better match of the extinction plasmon with the exciting laser. Moreover, the larger SERS intensity obtained for RC can be explained on the basis of the higher resonance effect obtained for this dye at the 532 nm excitation.

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Oral Multi-purpose X-ray diffractometer platform with versatile SAXS/WAXS options

Jan Gertenbach 1 J. Bolze 2 V. Kogan D. Beckers1 1 P. Munk and 1 M. Fransen

PANalytical B.V., 7602 EA Almelo, The Netherlands DANNALAB B.V., 7543 BK Enschede, The Netherlands

jan.gertenbach@panalytical.com

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A multi-purpose laboratory X-ray diffractometer was configured for advanced small angle X-ray scattering (SAXS) and wide angle X-ray scattering (WAXS) measurements. The modular concept of the instrument incorporates pre-aligned optical components and sample stages and thus allows for a quick and easy reconfiguration for a wide variety of applications. The high-end, user-friendly SAXS/WAXS setup is compact and includes an evacuated beam path. It enables measurements in an angular range of 0.08 – 78° 2θ and is also suitable for weakly scattering, dilute samples. Basic 2-dimensional (2D) SAXS experiments can also be undertaken. The EasySAXS software offers a user-friendly, comprehensive toolbox for data analysis.

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Oral Design of an efficient Graphene-based TiO2 nano-composite for photocatalytic removal of pharmaceuticals from water

Zahra Gholamvand 2 K. Nolan 1 J. Tobin A. Morrissey3

zahra.gholamvand@dcu.ie 1

School of Biotechnology, Dublin City University, Dublin 9, Ireland. School of Chemical Sciences, Dublin City University 3 Oscail, Dublin City University, Dublin 9, Ireland, Dublin 9, Ireland

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effect of chemical bonding between TiO2 and graphene, all composites were prepared by in-situ synthesis of TiO2 in the solution on graphene and also physical mixing of pre-synthesised bare TiO2 and graphene. The composite materials were systematically evaluated for the degradation of Diclofenac, Carbamazepine and Famotidine under both UV and visible light irradiation in a designed immersion well reactor shown in figure 1(f). Results and Conclusions In-situ synthesis of TiO2 nanostructures on GO yields uniform composite with strong physical and chemical binding without any free particle (figure 2(a),(c)(d)) which enhances the photocatalytic efficiency when compared to simple solution mixing and pure TiO2 (figure 1b). In natural pH TiO2 forms solid fibres during the hydrothermal process which grow uniformly on the surface of GO with ‎an average length of 1-2 micron and an average diameter of 10-20 nano meter (figure 1d). In highly alkaline solution, they form nanotubes on the surface of GO (figure 2c). ‎The highest photocatalytic performance under UV and visible was obtained for in-situ synthesised TiO2 nanotubes/GO composite. According to TEM, Raman spectroscopy, and BET analysis data, the superior photocatalytic enhancement was attributed to 1) the optimal assembly and interfacial coupling between the reduced GO sheets and TiO2 nanotubes, 2) the increase of surface-‎adsorbed reactant species due to large surface area (350 m2/g comparing 60 m2/g for P25) and meso-porous structure of nanotubes, 3) ‎the lower rate of photogenerated electron–hole pairs recombination due to the high conductivity of graphene, 4) the extension of the light ‎absorption range due to carbon doping in TiO2 during the hydrothermal process.‎ Almost 100% degradation of famotidine pollutant was achieved

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The presence of pharmaceutical compounds in the aquatic environment and their possible ‎effects on living organisms has emerged as a serious environmental concern particularly as conventional wastewater treatments only can partially remove them. The use of photocatalysts, particularly TiO2, has been considered one of the most cost and energy efficient clean technologies for use as tertiary wastewater treatment. The aim of this study is to find the most efficient TiO2/graphene nano-composite which is easy to process and able to purify water under solar light irradiation. The combination of TiO2 and reduced graphene oxide which ‎possess excellent adsorption, transparency, conductivity and controllability promises to simultaneously enhance pollutant adsorption and charge transportation to ‎facilitate photodegradation of the pollutants. Facile and reproducible routes to obtain chemically ‎bonded TiO2/graphene composites have been reported in the literature and their ability to degrade pharmaceuticals pollutants from water has been evaluated. The role of reduced graphene oxide platform in prohibiting electron-hole recombination and stimulating visible light activity of structurally controlled TiO2 photocatalyst (zero, one, and three-dimensional nano-structures shown in figure 1a) were also demonstrated. The effect of the dimensionality of TiO2 on photocatalytic performance, surface area, adsorption, and carrier transfer of the composites were investigated by preparing the homogeneous distributions of four different morphologies of TiO2 including nanoparticles, nanotubes, nanofibers and meso-porous beads via in-situ sol-gel method, alkaline hydrothermal method, solvothermal and sol-gel/solvothermal method respectively. This was followed by 2 h of post-thermal annealing at 400 C in Argon atmosphere. In order to study the

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after 40 min irradiation by UV and 100 min by visible light (figure 3-D1). In summary when ‎graphene is hybridized with TiO2, it can slow down the recombination of photo-‎generated electronhole pairs, increasing charge transfer rate of electrons and surface-adsorbed ‎amount of chemical molecules via π–π interactions which causes the enhancement in photocatalytic functionality of the composite [1].

References [1] P. V. Kamat. (2010 01/21). The Journal of Physical Chemistry Letters 1(2), pp. 520-527. Figures

Figure 1: of TiO2/graphene composite (a), the effect of synthesis method on photocatalytic efficiency (b).

Figure 2: SEM and TEM images of (a) sol-gel synthesised TiO2/GO, (b) solution mixing of P25/GO, (c): in-situ hydrothermally synthesised TiO2 nanotubes on RGO, (d): hydrothermally synthesised TiO2 nanofibres/GO, (e): 3D network of TiO2 beads/GO, (f): schematic image of immersion well Photocatalytic reactor.

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Figure 3: UV and visible light photocatalytic degradation of famotidine

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Invited Radiolabelling of Nanoparticles using Cyclotron-Based Techniques

Neil Gibson

neil.gibson@jrc.ec.europa.eu Institute for Health and Consumer Protection, Joint Research Centre, Ispra (VA) Italy

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Figure 1: The Scanditronix MC-40 Cyclotron at JRC Ispra, and the target capsule and holder used for direct ion-beam radiolabelling of dry nano-particulate powders.

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Radiolabelled nanoparticles can be used for tracing studies of relevance to many fields, such as nanomedicine, nanotoxicology, environmental tracing, occupational health, etc. Various methods for nanoparticle radiolabelling exist, each with advantages and disadvantages for particular studies. An outline is presented here of cyclotronbased techniques that can be used to directly radiolabel engineered nanoparticles, as well as some methods for synthesis of labelled nanoparticles using cyclotron-generated radioactive precursor materials. A basic outline is given of how cyclotron accelerators can be used to induce nuclear reactions, the characteristics of different radio-isotopes and their practical applicability for tracing, and what effect the direct ion-beam radiolabelling process might have on the nanoparticles themselves. In particular, the issues of thermal effects and radiation damage effects are addressed. Selected details are presented regarding some of techniques applied in our laboratories and those of collaborating groups, including direct light-ion activation of dry nanoparticle samples, neutron activation using a cyclotron ion-beam driven source, preparation of activated precursors and physical nanoparticle synthesis methods, recoillabelling via ion bombardment of mixed powders, and radiochemical synthesis of nanoparticles using radioactive precursors.

Invited New two-photon based nanoscopic modalities and optogenetics

DTU Fotonik, Dept. of Photonics Engineering, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark www.ppo.dk www.optorobotix.com

The science fiction inspired shrinking of macroscale robotic manipulation and handling down to the micro- and nano-scale regime open new doors for exploiting the forces and torques of light for micro- and nanobiologic probing, actuation and control [1-3]. A generic approach for optimizing light-matter interaction on these scales involves the combination of optimal light-sculpting [4] with the use of optimized shapes in micro-robotics structures [5]. Micro-fabrication processes such as two-photon photo-polymerization offer threedimensional resolutions for creating customdesigned monolithic microstructures that can be equipped with optical trapping handles for convenient mechanical control using only optical forces [6]. These microstructures illustrated below can be effectively handled with simultaneous top- and side-view on our BioPhotonics Workstation to undertake six-degree-of-freedom optical actuation of two-photon polymerised microstructures equipped with features easily entering the submicron-regime. Aided by European collaborators who fabricated test structures with built-in waveguides for us, we were able to put the idea of optically steerable freestanding waveguides – coined: wave-guided optical waveguides - to the test using our BioPhotonics Workstation [7]. We also propose using these techniques for generating two-photon real-time spatially sculpted light for the strongly emerging areas of neurophotonics and optogenetics [4].

[2] [3] [4]

[5]

[6]

[7]

Jesper Glückstad

jesper.gluckstad@fotonik.dtu.dk

microenvironments for biomedical studies," Optics Express 17, 6578-6583 (2009). J. Glückstad, “Sorting particles with light,” Nature Materials 3, 9-10 (2004). J. Glückstad, “Optical manipulation: Sculpting the object,” Nature Photonics 5, 7-8 (2011). E. Papagiakoumou, F. Anselmi, A. Begue, V. de Sars, J. Glückstad, E. Isacoff, V. Emiliani, “Scanless two-photon excitation of channelrhodopsin-2,” Nature Methods 7, 848-854 (2010). D. Palima, A. R. Bañas, G. Vizsnyiczai, L. Kelemen, P. Ormos, and J. Glückstad, "Wave-guided optical waveguides," Opt. Express 20, 2004-2014 (2012). D. Palima and J. Glückstad, “Gearing up for optical microrobotic manipulation: mechanical actuation of synthetic microstructures by optical forces,” Laser & Photonics Reviews (2013). H. Ulriksen, J. Thøgersen, S. Keiding, I. P.-Nielsen, J. Dam, D. Palima, H. Stapelfeldt, J. Glückstad, "Independent trapping, manipulation and characterization by an all-optical biophotonics workstation," J. Eur. Opt. Soc-Rapid 3, 08034 (2008).

Figures

References

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[1] P. Rodrigo, L. Kelemen, D. Palima, C. Alonzo, P. Ormos, and J. Glückstad, "Optical microassembly platform for constructing reconfigurable

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Invited Dielectric properties of biological samples measured at the nanoscale: from single bacteria to single viruses

Gabriel Gomila

gabriel.gomila@ub.edu Institut de Bioenginyeria de Catalunya (IBEC) and Universitat de Barcelona, Baldiri I Reixac 11-15, Barcelona (Spain)

[2] Gramse, G., Casuso, I., Toset, J., Fumagalli, L.

[3]

[4]

[5] [6]

[7]

and Gomila, G,. Nanotechnology 20, 395702 (2009). Fumagalli, L., Gramse, G., Esteban-Ferrer, D., Edwards, M. A. and Gomila, G., Appl. Phys. Lett. 96, 183107 (2010). Fumagalli, L., Esteban Ferrer, D., Cuervo, A., Carrascosa, J. and Gomila, G., Nature Materials, 11, 808–816 (2012). Fumagalli, L., Ferrari, G., Sampietro, M. and Gomila, G., Nano Lett. 9, 1604-1608 (2009). Gramse, G., Edwards, M., Fumagalli, L. and Gomila, G., Appl. Phys. Lett., 101, 213108 (2012). Gramse, G., Dols-Pérez, A., Edwards, M., Fumagalli, L. and Gomila, G., Biophysical Journal, 104, 1257-1262 (2013).

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Nanoscale dielectric microscopy (NDM) is a scanning probe microscopy technique that in addition to measure surface topography, enables to measure the local dielectric response (the dielectric constant, εr) of materials with nanoscale spatial resolution (< 50 nm). NDM can be performed in both AC current-sensing or forcedetection mode, and in recent years it has become a well applied technique for the study of dielectric materials [1-3]. Besides its application in materials science, it enables also to quantify the dielectric properties of biological samples and hence to quantitatively investigate electrostatic phenomena in biology. In the present communication, we will introduce nanoscale dielectric microscopy and present recent results obtained in the study of a variety of biological systems ranging from single viruses [4] and single bacteria to supported biomembranes [5]. In particular, we will show that the quantitative nature of the technique enables to develop a novel label-free material identification method based on the measurement of the dielectric response of the samples [4]. Examples include the identification of dielectric nanoparticles of different material composition [4], of single viruses from its empty capsids [4] or of single bacteria of different types. Finally, we will show how this technique can be extended to the liquid environment [6,7] opening a number of important possibilities in the study of electric processes in small scale biological systems References

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[1] Fumagalli, L., Ferrari, G., Sampietro, M. and Gomila, G. Appl. Phys. Lett. 91, 243110 (2007).

Oral Safety Implementation of Nanotechnology for Chemical Enterprises within a Bottom-Up Approach towards Communication

S. Hartl 1,2 F. Sinner Ch. Micheletti3 and A. Falk1

sonja.hartl@bionanonet.at

BioNanoNet Forschungsgesellschaft mbH, Graz, Austria 2 JOANNEUM RESEARCH- HEALTH, Graz, Austria 3 Veneto Nanotech ScpA, Padova, Italy This project is implemented through the CENTRAL EUROPE Programme co-financed by the ERDF.

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Within the aim of the Central Europe project NANOFORCE the general objective is to connect public and private organizations, to carry out collaborative and interdisciplinary researches on nanomaterials, and to turn the most promising laboratory results into innovative industrial applications. To build up a legal advisory board for chemical enterprises starting in nanotechnology, a state of the art report on existing safety procedures and nanotech related legislation was produced to illustrate the existing regulatory framework within the European region. Data sets of lab analysis which are performed on three different nanomaterials will lead to safety data sheets showing how the tested nanomaterials can be produced and professionally used conforming to safety and users guidelines. Results can already be shown generated by standardised in vitro testing methods for human toxicity and ecotoxicological testing for nanomaterials like nano-silver, zinc oxide and titanium dioxide. In the outline of presenting a transnational guideline on responsible management for researchers and industry, to have a safe and informative set of regulations to be followed when entering the nanotechnology businesses, another focus was laid on the correct implementation of nano-derived products and market placement, also partly focusing on consumer acceptance to be one of the major problems. Furthermore the guideline will help to distribute information on risks and benefits resulting from the use of nanotechnology as one of the key enabling technologies. Additionally the regions attractiveness for investors in nanotechnology within the CE area is presented by business plan incorporating information on micro-economic data, infrastructure and funding situation. This is needed

to foster the interactions between research and industry and additionally strengthened by a supervised online platform focusing on start-up collaborations between researchers and industry. This “NanoDeals- Generator” platform provides expertise tailored to individual needs and support innovative SMEs in launching new joint nanotechnology initiatives. Until now several experts have been registered and proposed their project ideas and NANOFORCE is providing a match-making facility to boost the communication activities. To evaluate the proposals a technology rating methodology for the assessment, validation and evaluation of “NanoDeals” has been developed following the “three-pillar bridge” presented by the High-Level Expert Group on Key Enabling Technologies, focusing on technology research, product demonstration and competitive manufacturing. Within this contribution and together with the current state of the art on regulations to be used for nanomaterials, necessities in regulation of nanomaterials will be presented. Preliminary test results in the field of nanosafety will be shown on different used and produced nanomaterials and directly provided by companies. Possible market barriers and including strategies to overcome these barriers are going to be presented within the NANOFORCE transnational guideline in terms of benefits resulting from nanotechnologies. In addition to this the “NanoDeals-Generator” online platform and the proposal of a business plan will be shown to outline the projects output - to fill the gap between more and less experienced regions and to turn investments in R&D into industrial innovations.

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Oral

Automated particle tracking (APT) for simultaneous zeta potential-, size distribution and concentration analysis of exosomes

Clemens Helmbrecht 2 K. de Miroschedji 2 A. -K. Ludwig B. Giebel2and 1 H. Wachernig

Particle Metrix GmbH, Neudiessener Str. 6, D-86911 Diessen, Germany Institute for Transfusion Medicine, University Hospital Essen, Hufelandstr. 55, 45122 Essen, Germany

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positions within the electro-optical cell to enhance statistical significance. As the particles are visualized, influences affecting the quality of the measurement such as agglomeration, fluctuation of concentration and background noise due to precipitation are better detectable by APT than with any other indirect characterization method. Particularly with regard to samples of low exosome concentration, the analysis of samples with particle concentrations as low as 0.5 million particles per mL can be performed. In summary, automated measurement of size, zeta potential and concentration of particles is the outcome of one simple measurement sequence. The analysis of several thousands of single particles is completed within minutes. Results on this technique will be demonstrated on selected exosome samples

References [1] Ludwig, A. K., Giebel, B., Exosomes: Small Vesicles Participating in Intercellular Communication, Int J Biochem Cell Biol. (2012), 44, 11-5. [2] Crocker, J. C., Grier, D. G., Methods of Digital Video Microscopy for Colloidal Studies, J. Colloid Interf Sci. (1996) 174, 298-310.

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Exosomes are extracellular vesicles between 30 and 170 nm in size range and occur naturally in bodyfluids. Research about the role of exosomes as transmitters of information between cells is a fast growing field which could open up the door to new approaches in diagnosis and therapy [1]. Research into this new field has been aggravated by the lack of techniques suitable for exosome characterization in buffers of high ionic strength. Direct 2D light scattering of liquid suspensions is an attractive instrument to assist researchers on the quantification of particle size, zeta potential and concentration. By tracking the 90째 scattering light of particles while undergoing Brownian migration, the particle size is derived by applying the Stokes - Einstein equation [2]. Trajectories of several thousands of single particles of the same sample are analyzed resulting in a representative particle size distribution (PSD) of high resolution (Fig. 1). The agglomeration of exosomes in the presence of the high ionic strength of the phosphate buffer (pH 6.5) resulted in a shift of the PSD towards larger hydrodynamic particle size. The electrokinetic properties of charged particles, i.e. electrophoretic mobility and zeta potential are determined by microelectrophoresis. In microelectrophoresis, the migration of particles in an electrical field is recorded. Electrophoretic mobility and zeta potential are calculated from the trajectory data by an automated algorithm. Fig. 2 shows examples of zeta potential distributions of exosomes determined in different buffers.The bench-top system is equipped with an automated start-up routine. After performing of auto-focus, auto-alignment and check of cell quality, the system is ready for measurements. Multiple measurements of concentration, particle size and zeta potential can be carried out in up to 11

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Figures

Figure 1: Effect of agglomeration of Exosomes isolated from BMBO cell line in phosphate buffer (pH 6.5) after 2 and 8 min of preparation.

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Figure 2: Zeta potential distributions of more than 400 single particles of each sample measured by automated particle tracking. Aquisition time was less than 2 min. actor.

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Oral Ultrafast laser spectroscopy of molecular tweezers

Wawrzyniec Kaszub 2 2 P. Uriac , B. Legouin 3 3 J. Graton , D. Jacquemin E. Hitti3and 1 M. Lorenc

IPR, UMR UR1-CNRS 6251, 263 av. Du General Leclerc, 35042 Rennes, France ISCR - UMR 6226, 2 Avenue du Pr. Leon Bernard CS 34317, 35043 Rennes, France CEISAM UMR CNRS 6230, Faculte des Sciences & des Techniques de Nantes 2, rue de la Houssiniere, BP 92208, 44322 Nantes, France 2 3

[6] J. Graton et al., J. Phys. Chem. (2012). [7] J. Graton et al., Chem. Phys. Lett. (2012).

Figures

Figure 1: Molecular tweezer synthetized with (1R,2R)-1,2diaminocyclohexane as spacer and two molecules of (+)-usnic acid as pincers [4].

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Femtosecond laser pulses open new perspectives to the ultrafast science, especially in that they allow controlling and observing molecular materials. Recent studies on molecular nanocrystals showed that pump-probe spectroscopy is an insightful method delivering information on the transient states. The same method was applied to the study of molecular tweezers, we present here. A molecular tweezer, which is essentially a hostguest pair of molecules, is a simple and efficient model to study and understand molecular recognition and self-assembly by Ď&#x20AC; - Ď&#x20AC; interaction. These non-covalent and hydrophobic interactions are involved in material science, catalysis and especially in biological processes and medical chemistry. A more recent and promising application of host-guest recognition is the design of molecular machines. In this context we have studied Pi0 and TNF complex with ultrafast laser spectroscopy, with the main goal being to reveal the mechanisms leading to the locking and unlocking of the guest by the host. The long term goal would be the laser control of such and related processes, and also design of functionalizable molecular tweezers.

maciej.lorenc@univ-rennes1.fr

References

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[1] R. Bertoni et al., Angew. Chem. Int. Ed. 51 (2012) 7458. [2] C. D'Amico et al., J. Phys. Chem. C 116 (2012) 3719. [3] M. Lorenc et al., Phys. Rev. Lett. 103 (2009) 028301. [4] B. Legouin et al. Org. Lett., 11, (2009) 745. [5] N. Ruangsupapichat et al., Nature Chemistry 3 (2011) 53.

Invited Sébastien Lecommandoux

Biomimetic polypeptide and polysaccharide based polymersomes for therapy and diagnosis

lecommandoux@enscbp.fr Université de Bordeaux, IPB-ENSCBP, 16 avenue Pey Berland, 33607 Pessac Cedex, France, CNRS, Laboratoire de Chimie des Polymères Organiques, UMR5629, Pessac, France

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Polymer vesicles (polymersomes) are among the most attractive systems for drug delivery applications. Actually, vesicles obtained by selfassembly of block copolymers are expected to overcome some of the current limitations in drug delivery, allowing the development of robust nanocontainers of either hydrophilic or hydrophobic species. In addition, the development of macromolecular nanodevices that can be used within the living body implies that sensors detecting chemical signals -such as ions, enzymes or pH changes- and generating internal signals or appropriate responses be integrated in the macromolecular system [1]. The use of peptide and saccharide building blocks in the copolymer structure would allow both controlling the selfassembled structure and the resultant biofunctionality. We report an overview on the self-assembly in water of amphiphilic block copolymers into polymersomes, and their applications in loading and controlled release of both hydrophilic and hydrophobic molecules and biomolecules. We pay special attention to polysaccharide and polypeptide-based block copolymer vesicles that we have studied these recent years in our group [2,3]. These newly developed copolymers that mimic the structure and function of glycoproteins represent an example of the effectiveness of a biomimetic strategy in implementing materials design [4]. In addition, magnetic polymersomes, including iron oxide γ-Fe2O3 nanoparticles are currently investigated, together with their potential applications as contrast agent for imaging and as therapeutic nanoparticles using hyperthermia [5]. Exciting and very promising results about their therapeutic evaluation for tumor targeting and in vivo tumor regression studies will be presented [6].

Finally our recent advances in using “biomimicry approaches” to design complex, compartmentalized materials will be proposed [7]. We demonstrate the formation of compartmentalized polymersomes with an internal «gelly» cavity using an original and versatile emulsion-centrifugation process. Such a system constitutes a first step towards the challenge of structural cell mimicry with both “organelles” and “cytoplasm mimics”. This study constitutes major progress in the field of structural biomimicry and will certainly enable the rise of new, highly interesting properties in the field of high-added value soft matter, especially in controlled cascade (bio) reactions

References [1] Garanger, E.; Lecommandoux, S. Angew. Chem. Int. Ed. 2012, 51, 3060-3062. [2] (a) Sanson, C.; Schatz, C.; Le Meins, JF.; Soum, A.; Thevenot, J.; Garanger, E. Lecommandoux, S. J. Control. Release (2010), 147, 428. (b) Sanson, C.; Schatz, C.; Le Meins, JF.; Brulet, A.; Soum, A.; Lecommandoux, S. Langmuir (2010), 26, 7953. (e) Sanson, C.; Le Meins, JF.; Schatz; Soum, A.; Lecommandoux, S. Soft Matter (2010), 6, 1722. [3] (a) Schatz, C.; Louguet, S.; Le Meins, JF.; Lecommandoux, S. Angew. Chem. Int. Ed. (2009) 48, 2572. (b) Upadhyay, K.K.; Le Meins, JF.; Misra, A.; Voisin, P.; Bouchaud, V.; Ibarboure, E.; Schatz, C.; Lecommandoux, S. Biomacromol. (2009), 10, 2802 [4] (a) Bonduelle, C.; Huang, J.; Ibarboure, E.; Heise, A.; Lecommandoux, S. Chem. Commun. 2012, 48, 8353. (b) Huang, J.; Bonduelle, C.;

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Thévenot, J.; Lecommandoux, S.; Heise, A. J. Am. Chem. Soc. 2012, 134, 119. [5] (a) Sanson, C.; Diou, O.; Thevenot, J.; Ibarboure, E.; Soum, A.; Brulet, A.; Miraux, S.; Thiaudiere, E.;Tan, S.; Brisson, A.; Dupuis, V.; Sandre, O.; Lecommandoux, S. ACS Nano (2011), 5, 1122. (b) Oliveira, H.; Pérez-Andrés, E.; Thevenot, J.; Sandre, O.; Berra, E. ; Lecommandoux, S. J. Control. Release (2013), ASAP (doi: 10.1016/j.jconrel.2013.01.013). [6] (a) Upadhyaya, KK.; Bhatt, A.N.; Mishra, A.N.; Dwarakanath, B. S.; Jain, S.; Schatz, C.; Le Meins, JF.; Farooque, A.; Chandraiah, G.; Jain, AK.; Misrac, AK.; Lecommandoux, S. Biomaterials (2010), 31, 2882. (b) Upadhyay, Mishra, AK.; Chuttani, K.; Kaul, A.; Schatz, C.; Le Meins, JF.; Misra, A.; Lecommandoux, S. Nanomedicine (2012), 8, 71. [7] (a) Marguet, M.; Edembe, L.; Lecommandoux, S. Angew. Chem. Int. Ed. 2012, 51, 1173. (b) Marguet, M.; Sandre, O.; Lecommandoux, S. Langmuir 2012, 28, 2035. (c) Marguet, M.; Bonduelle, C.; Lecommandoux, S. Chem. Soc. Rev. 2013, 42, 512-529.

Oral Cyclic measurements of DPPC monolayers at low surface tensions

Biolin Scientific, Tietäjäntie 2, FI-02130 Espoo, Finland

Pulmonary surfactants cover the alveoli of the lungs and have a vital function in making the process of breathing easy. During inhalation, the surfactant reduces the surface tension of tissue by a factor of about 15. During exhalation, the surface area of the alveoli decreases making the surfactant even more concentrated on the surface. It is known that the highly ordered solid phase of dipalmitoylphosphatidylcholine (DPPC) sustains the near-zero surface tension on the alveoli during exhalation [1]. In order to model the actual surfactant behavior in the alveoli, measurements at near-zero surface tensions are needed. Several groups have investigated the behavior of DPPC monolayers under low surface tensions using Langmuir troughs [2, 3], but with a conventional Langmuir trough it is challenging to measure nearzero surface tensions. We have shown that the compression speed in a Langmuir trough has a distinct effect on the layer formation of DPPC at low surface tensions and temperatures ranging from 20 ºC to 37 ºC. Now we further expand this observation by showing controlled cyclic measurements of DPPC at low surface tensions. The measurements were done on ultrapure water surface at temperatures of 20 ºC, 30 ºC and 37 ºC using a Langmuir trough equipped with a ribbon barrier to prevent monolayer leakage. The measurements show reliable compression measurements of natural phospholipid surfactants at surface tensions down to 15 mN/m. The measurements can be further expanded to examine phospholipid reactions with nanoparticles, phase transitions and the selective enrichment process of DPPC on the alveoli surface

Kaisa E. Lilja J. Välimaa

kaisa.lilja@biolinscientific.com

[3] Zhang, H.; Wang, Y. E.; Fan, Q.; Zuo, Y. Y. Langmuir 27 (2011) 8351.

References

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[1] Zuo, Y. Y. J. Appl. Physiol. 102 (2007) 1733. [2] Xicohtencatl-Cortes, J.; Mas-Oliva, J.; Castillo, R. J. Phys. Chem. B 108 (2004) 7307.

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Invited Biosensing with Gold Nanoparticles

Bionanoplasmonics Laboratory, CIC biomaGUNE, Paseo de Miramón 182, 20009 Donostia, Spain Ikerbasque, Basque Foundation for Science, 48011 Bilbao, Spain

llizmarzan@cicbiomagune.es

[3] R.A. Alvarez-Puebla, A. Agarwal, B.P. Khanal, P.

Aldeanueva-Potel, E. Carbó-Argibay, N. PazosPérez, E.R. Zubarev, N.A. Kotov, L.M. LizMarzán PNAS 108 (2011) 8157.

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Metal nanoparticles display very interesting optical properties, related to localized surface plasmon resonances (LSPR), which give rise to well-defined absorption and scattering peaks in the visible and near-IR spectral range. Such resonances can be tuned through the size and shape of the nanoparticles, but are also extremely sensitive towards dielectric changes in the near proximity of the particles surface. Therefore, metal nanoparticles have been proposed as ideal candidates for biosensing applications. Additionally, surface plasmon resonances are characterized by large electric fields at the surface, which are responsible for the so-called surface enhanced Raman scattering (SERS) effect, which has rendered Raman spectroscopy a powerful analytical technique that allows ultrasensitive chemical or biochemical analysis, since the Raman scattering cross sections can be enhanced up to 10 orders of magnitude, so that very small amounts of analyte can be detected. In this communication, we present several examples of novel strategies to employ colloidal nanostructures comprising gold or silver and silica in various morphologies, as substrates for ultrasensitive detection of a wide variety of analytes, including cancer markers, drug metabolytes and scrambled prions.

Luis M. Liz-Marzán

References [1] L. Rodríguez-Lorenzo, R. de la Rica, R. Alvarez-

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Puebla, L.M. Liz-Marzán, M.M. Stevens Nature Mater. 11 (2012) 604. [2] M. Sanles-Sobrido, L. Rodríguez-Lorenzo, S. Lorenzo-Abalde, A. González-Fernández, M.A. Correa-Duarte, R.A. Alvarez-Puebla, L.M. LizMarzán Nanoscale 1 (2009) 153.

Invited Skin applications of Atomic Force Microscopy

Gustavo S. Luengo

gluengo@rd.loreal.com L’Oréal Recherche and Innovation, Aulnay sous Bois, France

The skin surface can be observed with different methods more or less invasive. Atomic Force Microscopy (AFM) in vitro or ex-vivo allows a good resolution of the morphology of skin, not limited by the wavelength of light. In addition mechanical and chemical information is possible thanks to the physical interaction of a small nanometric probe on the surface. This presentation will show some of our recent studies on skin applications using this technology. In particular we will comment experimental aspects that are critical for performing this type of experiments from sample preparation to protocols of measurements (ambient, water, dry conditions, etc.). We will comment the specific morphological aspects like the micro topography of skin, the influence of age, the visualization of skin components. A special consideration will be given to the mechanical characterization of corneocytes, main constituents of the stratum corneum (outermost layer of skin). Their physical characterization from a elastic Young modulus determination to friction studies Finally we will show our current efforts to have access to chemical information by the use of modified AFM tips for protein recognition.

force microscopy. Experimental Dermatology 19 (11):1014-1019, 2010. [3] A. Potter, Gustavo S. Luengo, R. Santoprete and B. Querleux “Stratum Corneum Biomechanics” Ch. 16 in Skin moisturization V. Rawlings and J. J. Leyden Eds.., Informa Healthcare, 2009.

References [1] W. Tang and B. Bhushan. Adhesion, friction

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and wear characterization of skin and skin cream using atomic force microscope. Colloids Surf.B Biointerfaces 76 (1):1-15, 2010. [2] Christian Rankl, Rong Zhu, Gustavo S. Luengo, Mark Donovan, Nawel Baghdadli, and Peter Hinterdorfer. Detection of corneodesmosin on the surface of stratum corneum using atomic

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Oral Silica nanostructures toxicity assessment and their potential for biomedical applications

Maria Ada Malvindi

V. Brunetti1 G. Vecchio2 A. Galeone1 V. De Matteis1 R. Cingolani3 and P. P. Pompa1

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ROS. We found that SiO2NPs doped with iron oxide nanoparticles do not induce detectable cytotoxic effects up to 1 nM concentration (Fig.3b) with negatively charged NPs exerting the higher toxicity. This is likely associated to the nanoparticles degradation in lysosomal environment. Overall, we demonstrate that SiO2 nanostructures are quite safe in vitro and have promising potential in biomedical applications.

References [1] M.A. Malvindi et al., Nanoscale, 2012, 4; 4(2), 486-495. [2] G. Bardi et al., Biomaterials, 2010, 31, 65556566.

Figures

Figure 1: Representative TEM images of three sizes of SiO2NPs: 25, 60 and 115 nm.

Figure 2: a) Viability of A549 cells 48 and 96 h after the exposure to increasing doses evaluated of 25 nm SiO2NPs by the WST-8 assay; b) In vitro silencing of tGFP expression.

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Silica nanoparticles are widely used in various industrial fields and recently, they have been exploited also for biomedical research. The impact of SiO2NPs on human health and the environment is thus of great interest. Nowadays, the overall evaluation of the toxicity/biocompatibility of SiO2NPs is extremely difficult, owing to controversial results in the literature and to the lack of standard procedures and/or insufficient characterization of the nanomaterials in biological systems. Therefore the biocompatibility needs to be documented in greater detail. In this study we evaluated the toxicity of different silica nanostructures, both pure and quantum dots (QDs)- or iron oxide-doped, and studied their potential applications in gene delivery. We performed a systematic in vitro study to assess the biological impact of pure SiO2NPs, by investigating 3 different sizes (Fig.1) and 2 surface charges in 5 cell lines. We analyzed the cellular uptake and distribution of the NPs along with their possible effects on cell viability, membrane integrity and generation of reactive oxygen species (ROS). We observed that all the investigated SiO2NPs do not induce detectable cytotoxic effects (up to 2.5 nM concentration) in all cell lines (Fig.2a). Once having assessed the biocompatibility of SiO2NPs we evaluated their potential in gene delivery, showing their ability to bind, transport and release DNA, allowing the silencing of a specific protein expression (Fig.2b)1. The biocompatibility of SiO2NPs and their gene carrier performance were also evaluated and confirmed in primary neuronal cells2. Finally, we investigated the toxicity of silica nanoparticles doped with iron oxide nanocrystals. We tested nanoparticles with two surface charges in two cell lines by evaluating their effect on cell viability, cell membrane integrity and induction of

mariada.malvindi@iit.it

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Istituto Italiano di Tecnologia, Center for Bio-Molecular Nanotechnologies@Unile, Via Barsanti,1 â&#x20AC;&#x201C; 73010 Arnesano (Lecce), Italy Telephone number: +39 0832-1816248, Fax: +39 0832-1816230 2 Mawson Institute, University of South Australia, Mawson Lakes, SA 5095, Australia 3 Istituto Italiano di Tecnologia, Central Research Laboratories, Via Morego, 30 â&#x20AC;&#x201C; 16136 Genova, Italy

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Figure 3: a) SiO2NPs doped with iron oxide NPs; b) Viability of A549 cells 48 and 96 h after the exposure to increasing doses of SiO2NP doped with iron oxide NPs evaluated by the WST-8 assay; c) Iron release in lysosomal environment.

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Functionalization of silicon dioxide micropillars for biosensing and microarray technology

Lluis F. Marsal M. Alba P. Formentín P. Jacob J. Pallarès and J. Ferré-Borrull

Departament Departament d’Enginyeria Electrònica, Elèctrica i Automàtica, Universitat Rovira i Virgili, Avda Països Catalans 26, 43007 Tarragona (Spain)

lluis.marsal@urv.cat

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These micropillar arrays can be used as microarchitectured substrate for cell capturing and for biosensing. In all the cases the surface have to be functionalized in order to be selective to specific substances [7-8]. To demonstrate the feasibility of the chemical modification, the outer surface of the silicon dioxide micropillars was functionalized by silanization with 3aminopropyltriethoxysilane (APTES). Subsequently, the amine group of APTES is activated by incubation in glutaraldehyde (GTA). Finally, the samples were incubated with fluorescein-labelled bovine serum albumin (BSA-FITC). Figure shows a composed figure of scanning electron microscope (SEM) image of one inverted pyramid with micropillars and a schematic representation of the functionalization of the SiO2 micropillars. The FITC fluorescent labelling of the protein is used to confirm the BSA attachment onto the functionalized SiO2 micropillars. The observation under a fluorescence microscope revealed that the FITC-BSA bounds to the SiO2 micropillars. Figure 3 shows a confocal microscopy image top view of the silicon dioxide micropillars functionalized with APTES, GTA and FITC. Green colour comes from the FTIC-BSA functionalized surfaces. In conclusion, silicon dioxide micropillar arrays are potential biosensing platforms for molecular detection in biotechnological applications such as the detection of molecular binding events, substrates for cell culture and so on. Acknowledgements: This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO) under grant number TEC2012-34397, CONSOLIDER project CSD20070007 and by Generalitat of Catalunya under project 2009 SGR

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Macroporous silicon produced by electrochemical etching has been proved to be a promising material in a broad range of applications due to its versatility and low cost technology [1-3]. One interesting structure that can be obtained from macroporous silicon is silicon dioxide (SiO2) micropillar arrays. The uniformity and geometry of these micropillar arrays rely on the fabrication conditions as the micropillar structure is directly related with the pore geometry of the macroporous silicon. Briefly, these structures can be fabricated from macropores produced by electrochemical etching of n or p-type silicon wafers in hydrofluoric acid (HF) solutions. After the fabrication of macroporous silicon, the samples are thermally oxidized under oxygen atmosphere to obtain a silicon dioxide layer on all the surface. The last step is to remove the silicon dioxide layer on the backside of the wafer and to etch the silicon in TMAH solution to release the micropillars. The oxide within the macropores is resistant to the TMAH etch and silicon ioxide tips begin to appear on the backside of the wafer [4-5]. If the backside of the silicon wafer is patterned by lithographic techniques, we can obtain a platform of silicon dioxide pillar arrays inside truncated micropyramids. Figure 1 shows an example of silicon dioxide micropillar arrays inside of a set of truncated inverted pyramids. The size of the mask together with the etching time determine the dimensions of the pyramid. A large number of these structures can be fabricated on a single wafer and each one of those truncated pyramids can be used for detection of different molecular binding events [6] increasing at the same time the contact area between probes and target molecules due to the presence of the micropillars.

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Oral

References [1] V. Lehmann, Phys. Status Solidi (a), 197 (2003) 13. [2] V. Lehmann, Electrochemistry of Silicon:

[3]

[4]

[5]

[6] [7] [8]

Instrumentation, Science, Materials and Applications, Wiley-VCH, (2002). T. Trifonov, A. Rodríguez, L.F. Marsal, J. Pallarès, R. Alcubilla, Sensors and Actuators, APhysical 141 (2008) 662. T. Trifonov, A. Rodríguez, F. Servera, L.F. Marsal, J. Pallarès, R. Alcubilla, Phys. Status Solidi (a), 202 (2005) 1634. A. Rodríguez, D. Molinero, E. Valera, T. Trifonov, L.F. Marsal, J. Pallarès, R. Alcubilla, Sensors and Actuators, B-Chemical 109 (2005) 135. V. Lehmann, Nature Materials 1, (2002) 12. M.J. Sweetman, C.J. Shearer, J.G. Shapter, N.H. Voelcker, Langmuir, 27 (2011) 9497. I. Mey, C. Steinem, A. Janshoff, Journal of Materials Chemistry, 22 (2012) 19348.

Figure 3: Confocal microscopy top view of functionalized and fluorescent labelled (APTES + FITC) a) ordered SiO2 micropillars and b) single unattached SiO2 micropillar. The green color indicates the fluorescence of FTIC.

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Figure 1: SEM image of a set of inverted truncated pyramids with disordered SiO2 micropillars.

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Figure 2: SEM image of an inverted truncated pyramid with disordered SiO2 micropillars and schematic functionalization of micropillar.10 μm.

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Oral KTP nanocrystals exhibiting high SHG emission for bio-imaging applications 1

Laboratoire de Physique de la Matière Condensée – Ecole Polytechnique-CNRS, UMR 7643, 91128 Palaiseau Cedex, France 2 Laboratoire de Photonique Quantique et Moléculaire – ENS Cachan & CNRS, UMR 8537, 94235 Cachan, France 3 CEA/CNG, Laboratoire Génomique Fonctionnelle, 91057 Evry, France 4 INSERM 894, Centre Psychiatrie & Neurosciences de l’INSERM, 75014 Paris, France

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A. Slablab2 G. Dantelle1, V. Jacques2 A.-M. Lepagnol-Bestel3 S. Perruchas1, P. Spinicelli2 A. Thomas4, D. Chauvat2 M. Simonneau4 T. Gacoin1 and J.-F. Roch2

ludovic.mayer@polytechnique.edu thierry.gacoin@polytechnique.edu jean-francois.roch@ens-cachan.fr

References [1] F. C. Zumsteg, J. D. Bierlein, T. E. Gier, J. Appl. Phys. 47, (1976) 4980. [2] L. Le Xuan, C. Zhou, A. Slablab, D. Chauvat, C.

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Tard, S. Perruchas, T. Gacoin, P. Villeval, J.F.Roch, Small, 4(9) (2008) 1332.

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In the last few decades, numerous works have been devoted to the development of efficient luminescent probes for bio-imaging applications. KTiOPO4 (KTP), a well-known material for its nonlinear optical properties (especially second harmonic generation) [1] could be a good candidate as nonlinear optical probe because of its SHG excitation windows laying in the near IR range (transparent for biological tissues) and the possibility to determine the crystal orientation allowing orientation tracking [2 ]. We present here the coprecipitation synthesis of single-crystal KTP nanoparticles whose size ranges from 200 nm to less than 30 nm. They can be easily dispersed in water and are stable at physiological pH. Using a pulsed Ti-Sapphire laser at a wavelength of 1020 nm and a standard twophoton microscope combined with an Atomic Force Microscope, non-linear optical properties of these particles are investigated. They exhibit a perfectly stable high SHG signal and their crystalline orientation can be fully determined using the SHG high sensitivity to the polarization of the excitation light. We also show the possibility to use these particles as nonlinear optical probe for biology by introducing KTP nanoparticles into a culture medium with mouse primal neural cells. Investigations prove that individual KTP nanoparticles can be imaged inside the cell body and the neurites, while the absence of any visible impact on the short term development of the cells indicates their low toxicity at least for our in-vitro experiments.

Ludovic Mayer

Invited Multiple-probe scanning probe microscopy: a potential application to bio-inspired materials research

WPI Center for Materials Nanoarchitectonics (MANA), NIMS, 1-1 Namiki, Tsukuba, Ibaraki, Japan 2 Grad. School of Pure and Applied Sciences, Univ. of Tsukuba, 1-1 Namiki, Tsukuba, Ibaraki, Japan

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Multiple-probe scanning tunneling microscope (MP-STM) [1] enables us to investigate, at the nanometer scale, electrical properties of nanomaterials and functions of extended nanosystems as schematically illustrated in Figure 1. So far, MP-STM has been developed and used for characterizing electrical properties of nanomaterials, such as fullerene polymers, selforganized metal silicide nanowires, carbon nanotubes and so on [2]. However, our recent development to convert the MP-STM into MPatomic force microscope (MP-AFM) [3] really opens possibilities to perform MP-scanning probe microscope (MP-SPM) measurements on a variety of materials, ranging from metal, semiconductor, insulator and even biomaterials. One of the challenging and exciting research which can be done using MP-SPM would be the application of MP-SPM to biology. Since MP-SPM is, in principle, a device equipped with multiple inputs and outputs to/from materials and systems of interest, application of MP-SPM, for example, to a single cell would establish a signal processing cell system. Here, an important thing is to use appropriate probes for detecting signals in biosystems, i.e., molecular signals. Therefore, we have developed a tungsten suboxide probe which greatly enhances Raman scattering from molecules in a solution. Another example of interesting research is a development of neuromorphic architecture for neural network computation with MP-SPM. One of the major tackles in creating neural network based on silicon technology lies on the difficulty in preparing highly cross-over interconnections between units (some equivalent circuits to mimic neuron cells and synapses). Recently, an inorganic metal compound system was found to form neuromorphic structures [4]. We discuss how our

Tomonobu Nakayama1,2 1 J. Xu 1 R. Creasey 1 Y. Shingaya and 1 M. Aono

nakayama.tomonobu@nims.go.jp

MP-SPM can contribute to the development of neuromorphic network research toward future computation, together with preliminary results in forming bio-inspired neuromorphic network using carbon-based nanomaterials.

References [1] M. Aono et al., OYO Buturi 67 (1998) 1361. (in Japanese). [2] T. Nakayama et al., Advanced Materials 24 (2012) 1675. [3] S. Higuchi, O. Kubo, H. Kuramochi, M. Aono

and T. Nakayama, Nanotechnology 22 (2011) 285205. [4] A.Stieg, A. Avizienis, H. Sillin, C. Martin-Olmos, M. Aono and J. Gimzewski, 24 (2012) 286.

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Figure 1: Possible measurements using scanning tunneling microscopes with (a) single, (b) double, (c) triple, and (d) quadruple probes.

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Invited Structural characterization and mimicking of biological nanostructures in native heterogeneous environment

Niels Chr. Nielsen

ncn@inano.au.dk

Interdisciplinary Nanoscience Center (iNANO) and Department of Chemistry, Aarhus University, Aarhus, Denmark

[3] S. Zhang, M. Andreasen, J. T. Nielsen, L. Lei, E.

H. Nielsen, T. Skrydstrup, F. Besenbacher, N. C. Nielsen, D. E. Otzen, and M. Dong, Proc. Natl. Acad. Sci. USA, in press (2013). [4] N. V. Kulminskaya, M. Ø. Pedersen, M. Bjerring, J. Underhaug, M. Miller, N.-U. Frigaard, J. T. Nielsen, and N. C. Nielsen, Angew. Chem. Intl. Ed. 51 (2012) 6891. [5] C. U. Hjørringgaard, B. S. Vad, V. V. Matchkov, S. B. Nielsen, T. Vosegaard, N. C. Nielsen, D. E. Otzen, and T. Skrydstrup, J. Phys. Chem. B 116 (2012) 7652. [6] M. N. Burkhardt, R. Taaning, N. C. Nielsen, and T. Skrydstrup, J. Org. Chem. 77 (2012) 5357.

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In the quest for obtaining inspiration from Nature's advanced nano machineries, it is of great interest to establish high-resolution structure information of biological macromolecules in their heterogeneous, native, and functional environment. This is not trivial using traditional high-resolution methods such as X-ray diffraction and liquid-state NMR spectroscopy requiring the molecular entities either being in wellordered 3D crystals or in a state with fast molecular tumbling in solution. With examples addressing antimicrobial peptides [1], amyloid fibrils [2,3], and the photo antenna system of the chlorosomes in green sulfur bacteria [4], we demonstrate that combinations of solid- and liquidstate NMR spectroscopy, AFM/SPM, SAXS, TEM, cryo-EM, and MRI offers great potential to explore structure and dynamics of proteins residing in functional, heterogeneous environment. In addition to providing fundamental biological insight, these studies motivates the design of artificial nanoscale ion channels (Fig. 1) [5], biomarkers for early-stage detection of dementia [6], and artificial photo receptors.

References [1] J. Dittmer, L. Thøgersen, J. Underhaug, K.

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Figure 1: Artificial antimicrobial ion channel.

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Bertelsen, T. Vosegaard, J. M. Pedersen, B. Schiøtt, E. Tajkhorshid, T. Skrydstrup, and N. C. Nielsen, J. Phys. Chem. B 113 (2009) 6928. [2] J. T. Nielsen, M. Bjerring, M. D. Jeppesen, R. O. Pedersen, J. M. Pedersen, K. L. Hein, T. Vosegaard, T. Skrydstrup, D. E. Otzen, and N. C. Nielsen, Angew. Chemie Intl. Ed., 48 (2009) 2118, P. Villeval, J.-F.Roch, Small, 4(9) (2008) 1332.

Oral Multifunctional Coordination Polymer Particles for Bio-medical application

Fernando Novio 1 P. González-Monje 2 J. Lorenzo and D. Ruiz-Molina1

Centro de Investigación en Nanociencia y Nanotecnología, Campus UAB, 08193 Bellaterra (Spain) 2 Unitat d´Enginyeria de Proteïnes i Proteòmica. Institut de Biotecnologia i Biomedicina Universitat Autònoma de Barcelona, 08193 Bellaterra (Spain)

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The development of different techniques to achieve nanomaterials with unusual and enhanced properties compared to traditional materials have afford new systems with interesting applications in areas such as electronics, catalysis, sensing or biomedicine. The coordination polymers are a fascinating family of materials created from supramolecular assembly of metal ions or clusters and organic ligands that act as building blocks to generate a superstructure with genuine and highly tailorable properties. The results obtained in the last years have shown that these systems can be interesting alternative materials for technological uses. The easy of tuning and modification of their properties based on chemical-structural changes allow control rationally their physicochemical properties. Our principal efford within our research group is to develop an specific kind of coordination polymers particles synthesized at the nanoscale with application in Biomedicine.[1] In this research area there is an special interest in achieve multifunctional platforms that can provide different properties with interest in biomedicine.[2] A posible ideal system would consist in a nano-platform with optimun size and shape to penetrate in different tisues but also to avoid the immunological system, that is be able to encapsulate different active substances, with biosensing properties and active as bioimaging agent. Moreover, the posibility of building these systems containing the active specie as buildingblock open a new approximation to achieve a platform with smart activity and added value. With this aim, our research group has developed the synthesis of nanoscale polymeric coordination particles (CPPs)1 able to encapsulate a wide variety of substances and materials for biomedical

fnovio@cin2.es

application.[3,4] The last experimental results have afford the synthesis and characterization of smart responsive CCPs able to tune such release upon external stimuli.[5] We will also show our most recent advances for their use not only for delivery but also for bioimaging purposes as well as the possiblity to functionalize them to improve their cell internalization processes, biocompatibility, and targeting directionality for especific recognition.[6] The good physicochemical properties of these nanoplatforms joint to the extremelly low toxicity, make them an interesting alternative to the current systems. Here we report the synthesis and characterization of CPPs with controllable dimensions (between 40nm-2µm) bearing reactive groups (-COOH, -NH2, etc.) at the surface. Their crystallinity and morphology can be controlled systematically by the proper combination of ligands and metal ions and reaction conditions. Using carbodiimide assisted coupling reactions different active molecules and biomolecules have been attached to the nanoparticles surface confering them a given specific property. In that manner we have been able to graft CPPs with dyes or poly-ethylene glycol derivates, among others. These ligands confer new interesting properties such as fluorescene to follow the cellular internalization of the CPPs or robustness against opsonization processes. As complementary studies, we have achieved the robust immobilization of these functionalized systems in gold surfaces. This approximation provides interesting platforms to be used in integrate systems with potential uses in biosensing and in clinical diagnosis All these preliminary results joint to the cytotoxicity experiments that shown a very good

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biocompatibility of the resulting functional capsules even for the case of using cobalt metal ions, open new venues for CPPs to be used as multifunctional platforms at biological level.

References [1] . Novio, J. Simmchen, N. Vazquez, L. Amorín,

[3]

[4]

[5]

[6]

Figure 2: a-b) TEM image of iron oxide nanoparticles, c) interaction of the loaded nanoparticles with a magneto, and d) hysteresis loop of encapsulated magnetic nanoparticles

Figure 3: Scheme of fluorescein functionalized CCPs by carbodiimine assisted coupling reaction.

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[2]

D. Ruiz-Molina, Coord. Chem. Rev. submitted invited paper. M. De, P. S. Ghosh, V. M. Rotello Adv. Mater. 20, (2008), 4225. I. Imaz, J. Hernando, D. Ruiz-Molina, D. Maspoch, Angew. Chem. Int. Ed. 48, (2009), 2325. I. Imaz, M. Rubio-Martínez, L. GarcíaFernández, F. García, D. Ruiz-Molina, J. Hernando, V. Puntes, D. Maspoch, Chem. Commun., 46, (2010), 4737. F. Novio, J. Simmchen, J. Ruiz, V. Rodriguez, N. Cutillas, J. Lorenzo, D. Ruiz-Molina, paper in preparation. F. Novio, P. González-Monje, J. Lorenzo, D. Ruiz-Molina, Angew. Chem. Int. Ed. Engl. Submitted.

Figure 4: Optical and fluorescence images of the fluorescent nanoparticles internalization.

Figure 1: a) Scheme of CPPs synthesis, b) SEM images of the resulting nanoparticles, and c) TEM image of a drug loaded nanoparticles.

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Figure 5: HR-SEM images of CPPs attached to gold surface.

Invited Singlet Oxygen in Mammalian Cells: Exploiting Nanoscopic Tools in Single Cell Experiments

Peter R. Ogilby

progilby@chem.au.dk

Center for Oxygen Microscopy and Imaging, Department of Chemistry, Aarhus University, Aarhus, DK-8000, Denmark

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Singlet oxygen, the lowest excited electronic state of molecular oxygen, is a â&#x20AC;&#x153;mature citizenâ&#x20AC;? that has been studied for many years from a wide range of perspectives. Among other things, singlet oxygen has a unique chemistry that results in the oxygenation of many organic molecules. In this way, it plays important roles in biology, particularly in mechanisms of cell signaling and cell death. Singlet oxygen is commonly produced in a photosensitized process wherein light is absorbed by a given molecule (the so-called sensitizer) followed by energy transfer from the excited state sensitizer to ground state oxygen. The production of singlet oxygen in this way is a natural phenomenon (i.e., we live in a world of light, oxygen, and suitable sensitizers). We have a multi-faceted program in which the behavior of singlet oxygen is examined under conditions and in systems that have, heretofore, been inaccessible. We are particularly interested in developing and exploiting tools that provide unique insight into the mechanistic behavior of singlet oxygen in cells. I will briefly describe our latest work that is relevant to phenomena on the nano-scale: (a) focused two-photon sensitizer excitation to impart spatial selectivity in single cell experiments, (b) protein-encapsulated sensitizers that can be localized in specific intracellular domains, (c) electric fields associated with nanoparticle surface plasmons to enhance radiative transitions in oxygen, and (d) added quenchers to influence diffusion-dependent singlet oxygen deactivation. Our results indicate that there is still much to be gained from studies of singlet oxygen.

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Invited Enzymes and fluorescent nanoparticles

Valeri Pavlov L. Saa and G. Garrai

vpavlov@cicbiomagune.es CIC BiomaGUNE, Paseo Miramon 182, San Sebastian, Spain

References [1] Saa L., Virel A., Sanchez-Lopez J., Pavlov V., Chem-Eur. J. 16 (2010) 6187. [2] Saa L., Pavlov V., Small, 8 (2012) 3449.

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Semiconductor inorganic NPs can be photoexcited to generate electron/hole couples which recombine to yield fluorescent emission of light. The stable and bright emission of semiconductor NPs arises from quantum confinement effects that occur in nanometer-sized semiconductors hence such NPs are called quantum dots (QDs). QDs modified with antibodies, DNA and small molecules found application as labels in bioanalytical affinity assays and imaging. Decorated hybrid NPs, iron oxide NPs and nanostructures with an elongated assembly of cores are utilized in magnetic resonance and fluorescence imaging for detection of cancer. The above mentioned analytical systems are based on presynthesized NPs decorated with recognition elements acting as labels or donor/quencher FRET pairs for quantification of enzymatic activities. Their performance is frequently hindered by high background signals caused by nonspecific adsorption of decorated NPs on surfaces or poor quenching of donor couples. Generation of NPs in situ can address these drawbacks of relevant analytical systems by decreasing the background signals. We developed some methods employing enzymatically driven formation of CdS QDs. The examples are represented in Scheme 1, Scheme 2, Scheme 3 and Scheme 4. The above mentioned enzymatic assays demonstrated sensitivity better by one-two orders of magnitude than previously reported assays using commercially available chromogenic substrates

Scheme 1: Assay for acetylchololinesterase (AChE)1.

Scheme 2: Assay for alkalinephosphatase (ALP)1.

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Scheme 3: Assay for glucose oxidase (GOx)2.

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Scheme 4: Assay for horseradish peroxidase (HRP)2.

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Invited Molecular nanomechanics in human health

Raul Perez-Jimenez

rperezjimenez@nanogune.eu CIC nanoGUNE Tolosa Hiribidea 76, Donostia-San Sebastian, Spain

[2] Wiita AP, Perez-Jimenez R et al. Nature, 450(7166), 2007, pp 124-127. [3] Perez-Jimenez R et al. Nat Struct Mol Biol,

16(8), 2009, pp 890-6.

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Mechanical forces play a crucial role in a myriad of biological processes including numerous diseases and disorders [1]. However, molecular nanobiomechanics are barely considered in modern medicine. Our research line focuses on atomic force microscopy (AFM) to investigate the effect of mechanical forces in proteins and enzymatic reactions that are relevant to human pathologies. In the past years, we have investigated how force affects the chemistry of thioredoxins, a class of enzymes that regulate the redox balance in cells. We showed that force regulates their chemistry, revealing several mechanisms for disulfide bond reduction that were hidden in standard bulk assays [2,3]. Importantly, these enzymes seem to be important during HIV-1 infection by reducing disulfide bonds in human CD4, the primary receptor of the virus. Using AFM techniques we have investigated the mechanics of CD4. We observed that force might trigger mechanical unfolding of CD4 and subsequent disulfide bond reduction in CD4 by Trx enzymes. Further experiments demonstrated, for the first time, that an antibody that blocks HIV-1 infection produces a mechanical effect on CD4 by preventing mechanical unfolding. We suggest that mechanical force might be important during HIV-1 infection which may change our understanding of the mechanism of infection. This observation might offer new avenues to explore novel treatments focused in the mechanics of proteins and enzymes, that is, mechanopharmacology.

References

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[1] Vogel V, Sheetz MP, Nanomedicine. WileyVCH, Wenheim, (2009) pp 235-303.

Oral Multifunctional Nanoplatform for Biomedical Applications 1

Rafael Piñol

Ins. de Ciencia de Materiales de Aragón (ICMA), CSIC–Univ. of Zaragoza, C/ Pedro Cerbuna 10, 50009 Zaragoza, Spain. 2 Departamento de Física and CICECO. University of Aveiro, 3810-193 Aveiro, Portugal. 3 Lab. of Molecular Toxicology, Fac. of Veterinary Medicine, Univ. of Zaragoza, 50009 Zaragoza, Spain. 4 Department of Hematology, Faculty of Medicine, University of Zaragoza, 50009 Zaragoza, Spain. 5 Department of Physics “A. Volta”, University of Pavia, 27100 Pavia, Italy. 6 Laboratoire de Physique et Chimie des Nano-Objets (LPCNO), INSA-University of Toulouse, 135, av. de Rangueil, F-31077 Toulouse, France. 7 Ins. de Química Avanzada de Cataluña (IQAC), CSIC, Jordi Girona, 18-26, 08034, Barcelona, Spain. 8 Centro de Investigación del Cáncer (CIC), CSIC-Univ. of Salamanca, 37007 Salamanca, Spain.

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After a period of intense research, nanoparticles incorporating a biological functionality have already found several punctual applications in biotechnological and clinical practices. Nowadays, the development of functionalization techniques with a high versatility permits to aspire to a more general use of nanoparticles in nanomedicine. In this direction, the concept behind this work pretends to use nanoparticles as a platform for assembling multiple interchangeable pieces each of them incorporating a physical or biological functionality. The nanoplatform presented here shows that it is possible to combine physical, chemical and biological functionalities in a single particle and simultaneously. It consists on a hydrophobic polymer core having a surface with multiple equal anchoring groups for the covalent bonding of the functionalised pieces. This core in itself may also incorporate several functionalities in the form of smaller inorganic nanoparticles. The functionalised pieces consists of hydrophilic chains ending on a reactive group that fits on the core surface and holds at the other end a biological or physical functionality. The coupling system is a Michael addition from a Michael-donor (amine, thiol, etc) to a Michael acceptor (acrylate) [1]. This system has the advantages of a clean synthesis (no by-products), mild conditions, and an easy and controlled multifunctionalization. The nanoplatform has been functionalized with radiochemical tracers (In111), luminescent dyes (fluorescein, rhodamine, lanthanide compounds), and magnetic nanoparticles, and therefore it can be a powerful tool in imaging. Besides, it has also been functionalized with a therapeutic drug, an antibody, and an optical thermometer made of lanthanide complexes [2]. Health safety of the system has been tested in cellular and in vivo assays. The nanoplatform is highly stable in

L.M.A. Ali1, R. Bustamante1 C. Brites2, L. Gabilondo1, J.L. Murillo1, N.J.O. Silva2 V. Sorribas3, M. Gutierrez4 R. Cornudella4, L. Carlos2, F. Palacio1, A. Lascialfiari5 J. Carrey6, M. Respaud6 G. C. Sanmartí7, J.-P. Salvador7 M.-P. Marco7, J. Criado8 M. Fuentes8 and A. Millan1

pinol@unizar.es amillan@unizar.es palacio@unizar.es

biological fluids, shows low cell toxicity [3], high capacity of cell internalization, no in vivo damage for tissues and organs, excellent hematocompatibility, and anticoagulation properties [4]. It is shown that magnetic properties can be tuned up in the whole superparamagnetic range [5]. Moreover, the system has shown excellent performance in magnetic resonance imaging [6] and hyperthermia

References [1] R. Piñol, A. Millán, F. Palacio, L. Gabilondo,

[2]

[3]

[4]

[5]

[6]

Spanish Patent no. P201031493 submitted to the Spanish Patent and Trademark Office (0710-2010). C. D. S. Brites, P. P. Lima, N. J. O. Silva, A. Millán, V. S. Amaral, F. Palacio, L. D. Carlos, Advanced Materials, 22 (2010) 4499-4504. R.Villa-Bellosta, G. Ibarz, A. Millan, R. Piñol, A. Ferrer-Dufol, F. Palacio, V. Sorribas, Toxicology Letters, 180, (2008) S221-S2216. Lamiaa M.A. Ali, M. Gutiérrez, R. Cornudella, J.A. Moreno, R. Piñol, L. Gabilondo, A. Millán and F. Palacio, J. Biomed. Nanotechn. (in press) (2013). A. Millan, F. Palacio, A. Falqui, E. Snoeck, V. Serin, A. Bhattacharjee, V. Ksenofontov, P. Gütlich and I. Gilbert, Acta Materialia, 55, (2007), 2201-2209. H. Amiri, P. Arosio, M. Corti, A. Lascialfari, R. Bustamante, A. Millán, N.J.O. Silva and F. Palacio, Magn. Res. in Medicine, 66, (2011) 1715-1721.

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Figure 1: Hydrophobic polymer Fluorescent probe Magnetic core (ÎłFe2O3) Hydrophilic polymer (PEG) Hydrophilic polymer with anchoring groups (PEG) Antibody Scheme of core-shell multifunctional nanoplatform containing magnetic nanoparticles.

Oral Nanoparticles of plant origin exploited for multiple applications

Fernando Ponz F. Sรกnchez, C. Mansilla P. Ibort, S. Cuenca M. Aguado, C. Cruz and I. Gonzรกlez.

CBGP (UPM-INIA). Montegancedo, Pozuelo, Madrid, Spain

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Plant-made viral nanoparticles (VNPs) are natural nano-objects with the characteristics of typical nanoparticles such as size, monodispersity and symmetry, which can be exploited as scaffolds for building modified nanostructures with multiple applications. In addition, plant VNPs are produced at a rather low cost; being composed of proteins they are completely biocompatible; and they do not have the ability to infect animals or humans, thus offering an extremely high degree of biosafety. (VNPs) can take several forms, which mainly fall into two biological categories. Either the particles enclose and pack a nucleic acid (virions) or they do not (virus-like particles, VLPs). Each of them has its particular pros and cons for nanotechnological deployment. The modifications performed on VNPs also can be achieved by two main approaches: genetic or chemical modifications. Using combinations of these possibilities we are currently developing VNPs derived from Turnip mosaic virus (TuMV) as a nanoplatform for several applications. Each TuMV VNP is a flexuous nanorod with a diameter of ca. 15nm, made up of over 2000 identical protein subunits helically arrayed (Figure 1). This structure can be conveniently modified for different purposes depending on the particular application sought. We will be presenting and discussing our progress in the exploitation of TuMV VNPs as scaffolds for peptide and protein presentation. These modifications have allowed us to develop applications in the areas of enhanced immunization and antibody production, sensitive detection of antibodies and infectious agents, and enzyme nanoimmobilization for improved industrial biocatalysis.

Figures

Figure 1: Electron micrograph of purified TuMV VNPs, showing their structural characteristics of flexuous nanorods.

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Víctor Puntes J. Comenge, C. Sotelo F. Romero, O. Gallego A. Barnadas T. García-Caballero and F. Domíngue

Institut Català Nanotecnologia, Universidad Santiago de Compostela, Universidad de Lugo, Hospital de Sant Pau, Spain

victor.puntes@ic.cat

Last decade has seen a flourishment in the study of the properties of inogranic nanoparticles for medical applications. Nanoparticles display properties that are strongly determined by both morphology and environment and in the physicochemical context where they are immersed, therefore allowing to monitor and manipulate biological states. In fact, inorganic nanoparticles behave as "artificial atoms", since their high density of electronic states -which controls many physical properties- can be extensively and easily tuned by adjusting composition, size and shape and used in biological environments. In fact, nanotechnology’s ability to shape matter on the scale of molecules is opening the door to a new generation of diagnostics, imaging agents, and drugs for detecting and treating disease at its earliest stages. But perhaps more important, it is enabling researchers to combine a series of advances, creating thus nanosized particles that may contain drugs designed to kill tumours together with targeting compounds designed to home-in on malignancies, and imaging agents designed to light up even the earliest stage of cancers. In fact, a description of cancer in molecular terms seems increasingly likely to improve the ways in which human cancers are detected, classified, monitored, and (especially) treated, and for that, nanoparticles, which are small and therefore allows to address molecular structures in an unique manner, may be specially useful for those tasks. Thus, Nanoparticles (NPs) have emerged as a potential tool to improve cancer treatment. Among the proposed uses in imaging and therapy, their use as a drug delivery scaffold has been extensively highlighted. However, there are still some controversial points which need a deeper

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understanding before clinical application can occur. Here the use of gold nanoparticles (AuNPs) to detoxify the antitumoral agent cisplatin, linked to a nanoparticle via a pH-sensitive coordination bond for endosomal release, is presented. The NP conjugate design has important effects on pharmacokinetics, conjugate evolution and biodistribution and results in an absence of observed toxicity. Besides, AuNPs present unique opportunities as drug delivery scaffolds due to their size and surface tunability. Here we show that cisplatin-induced toxicity is clearly reduced without affecting the therapeutic benefits in mice models. The NPs not only act as carriers, but also protect the drug from deactivation by plasma proteins until conjugates are internalized in cells and cisplatin is released. Additionally, the possibility to track the drug (Pt) and vehicle (Au) separately as a function of organ and time enables a better understanding of how nanocarriers are processed by the organism.

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Detoxifying Antitumoral Drugs via Nanoconjugation: The Case of Gold Nanoparticles and Cisplatin

NANOBIO&MED 2013

Invited

Oral Nanofluidic system for matrix-free DNA analysis

Hubert Ranchon 2 J. Lacroix 1 C. BlatchĂŠ and A. Bancaud1

CNRS, LAAS; 7 avenue du colonel Roche, F-31400 Toulouse, FRANCE Univ de Toulouse, LAAS, F-31400 Toulouse, France

hranchon@laas.fr

354

Chromosomes are the universal support of the genomic material, and their analysis at the genome level is among the most active field of research and innovation. Most analytical tools have the ability to sequence short DNA fragments, which are subsequently registered by computers to define a complete genome. This cost-ineffective situation is mostly related to the history of molecular biology techniques, which have been developed to characterize DNA chains of less than ~20 kbp. We posit that micro- and nanotechnologies offer new solutions to manipulate long DNA fragments, and hold great promise to the analysis of large scale genomic rearrangements which are common in cancer [1]. In this report, we describe a novel technology to separate biomolecules by size, which does not require the use of a separation matrix. The system consists of a silicon fluidic chip obtained by conventional photolithography, followed by dry etching (Fig.1.a). The slit-like geometry enables to monitor the environment of biomolecules, and we use a combination of hydrodynamic and electrophoretic to convey biomolecules (Fig.1.b), so that three control parameters, namely the geometry, the pressure, and the electric field, enable to control the progression of biomolecules in the channels. The application of an hydrodynamic field alone does not induce separation, whereas the application of an electric field acting opposite to the hydrodynamic flow improves the performances of our system: the chromatogram obtained with 500 mbar and 30 V shows sharp peaks for ten DNA species spanning 500 to 10000 bp in less than 8 minutes (Fig. 2). Small DNA molecules migrate faster than large ones, as observed with conventional gel electrophoresis but not with hydrodynamic

separations [2]. Using real time single molecule nanoscale imaging, we show that size-dependent migration is achieved through transverse displacements: molecules accumulate along streamlines according to their molecular weight, so that their progression along channels is determined by their size. This phenomenon is enhanced with the geometry of the channel and with the rheological properties of the fluid, and we show the successful separation of DNA molecules from 100 bp to 100000 bp in minutes, a performance inaccessible to most technologies available on the market. Most importantly we demonstrate sharp variations of DNA velocity as a function of its size characterized by power-law scalings up to ~-2,75 (Fig. 3), thus outperforming conventional electrophoresis that is associated to a power-law scaling of -1 [3]. Overall this study is a demonstration of the potential of fluidic technologies for the life sciences. Our technology is readily adapted for integration in Lab-on-Chip applications, and it is well-suited to perform analytical operations on whole-chromosomes for novel biodiagnostic applications.

References [1] Stephens P.J. et al., Cell, 144 (2011) 27-40. [2] Liu K.J et al., Journal of the American Chemical Society, 133 (2011) 6898-6901. [3] Viovy J.L., Review of Modern Physics, 72

(2000) 813-872.

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Figures Buffer tanks RĂŠservoirs (solvant)

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Figure 1: Description of the integrated fluidic system and the x actuation technology. a) Sketch of the b) chip on a coverslip observed d) on an inverted microscope. b) All fluidicz tanks are pressured)and voltagez z Sideshow view controlled. b) Descriptionz of the actuation modes: red arrows x y line stands for the Poiseuille the electrophoretic field and the blue x y y flow. 3 kpb

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Figure 2: Chromatograms obtained for a commercial kb ladder @ [500 mbar; 30 V] demonstrating clear resolution of the 10 DNA strands of the sample in less than 8 minutes. Associated to each peaks are video frames obtained on the EMCCD camera, allowing for size discrimination based on single molecule intensity. The inset on the right is a photograph of the ladder after migration in an agarose gel. 2

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[200 mbar; 20 V] V_200_20V V_500_20_V [500 mbar; 20 V] V_500_30V [500 mbar; 30 V] V_700_30V [700 mbar; 30 V] V_700_40V [700 mbar; 40 V] Scaling_au_pif Scaling ~ bp -1.33

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Figure 3: Velocity of DNA molecules as function of their lengths. kb ladder (left). 100 bp ladder (right). Pressure drops and voltages are indicated in inset. Tuning of the electric field and pressure drop induces sharp variations in mobility with the molecular weight. The highest variations are associated to a power scaling of ~ -1.33 and ~ -2.75 (black dashed and dotted lines).

Oral Towards DNA-Coated Anisotropic Building-Blocks: A Robust Surface Modification Strategy to Functionalize Gold Nanorods with DNA Oligonucleotides

Jessica Rodríguez-Fernández V. Baumann and F. Haase

Jessica.Rodriguez@lmu.de

Ludwig-Maximilians-Universität (LMU), Amalienstr. 54, Munich, Germany

Anisotropic nanoparticles like gold nanorods (Au NRs) are appealing building-blocks for the development of higher-order assemblies with strong plasmon-plasmon interactions and thus, sensing potential. DNA-directed assembly is one of the most powerful and versatile strategies to controllably direct either the self-organization of colloidal nanoparticles [1] or the formation of biocompatible organic-inorganic nanoscale hybrids of complex geometries.[2] In this context, the effective functionalization of Au NRs with DNA is a necessary key step, where the challenge remains in avoiding NR aggregation throughout the DNAmodification process. In this work we have developed a strategy to functionalize CTAB-stabilized gold nanorods with DNA oligonucleotides while maintaining their colloidal stability.[3] Our approach consists of a ligand exchange process that leads to the careful displacement of CTAB from the Au surface by “thiolated” helper molecules first and by thiolated DNA strands in the last instance. We will present our results on the optimization of the different surface functionalization steps and on the quantification of DNA grafting on the Au NRs’ surface. We will also show that the asfunctionalized nanorods are highly stable overtime in high ionic strength media. Taken altogether, our results indicate that our DNA-functionalized Au NRs may be used as building-blocks for DNAdirected (self)-organization.

[2] J. K. Do, R. Schreiber., A. A. Lutich, T. Liedl, J.

Rodríguez-Fernández, J. Feldmann, Nano Letters, 12 (2012), 5008. [3] V. Baumann, F. Haase, J. Rodríguez-Fernández, in preparation.

References

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[1] C. A. Mirkin, R. L. Letsinger, R. C. Mucic, J. J. Storhoff, Nature, 382 (1996), 607.

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Oral Transformation and toxicity of PAMAM dendrimers under irradiation and ozonation processes

1,2

Roberto Rosal 2 J. Santiago-Morales 3 M. D. Hernando M. M. Ulaszewska1 1 E. García-Calvo and 1 A. R. Fernández-Alba

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corona discharge and continuously bubbled as described elsewhere [7]. Irradiation experiments were carried out using a 15W Heraeus Noblelight TNN 15/32 low-pressure mercury vapour lamp emitting at 254 nm and a Heraeus TQ Xe 150 Xearc lamp with spectral emission in the visible region. The temperature was 25°C and pH was controlled at 7.0 ± 0.1 units. Samples were withdrawn for analysis at prescribed intervals. Analytical. The analyses have been performed by liquid chromatography using a hybrid quadrupole/time of flight mass analyzer (LC-ESIQTOF-MS) and a TripleTOF 5600 System (AB SCIEX, Concord, ON) coupled to an HPLC Agilent 1200 (Agilent Technologies, Wilmington, DE, USA) with a ESI source. The AB SCIEX TripleTOF 5600 equipment offers the capability of high resolution detection with resolving power 40000 (FWHM). The LC analyses were performed with a reversedphase C5 analytical column 150 mm length x 4.6 mm I.D. and 5 µm particle size. The MS was operated in full scan TOF-MS and MS/MS mode through information dependent acquisition (IDA) in a single run analysis. The data acquisition and processing was carried out using Analyst® TF 1.5 and PeakViewTM. Toxicity. The toxicity of samples corresponding to partially oxidized and irradiated mixtures was assessed using the following bioassays: multigenerational growth inhibition of the green alga Pseudokirchneriella subcapitata (OECD TG 201), the inhibition of the constitutive luminescence of the marine bacterium Vibrio fischeri (ISO 11348-3 standard protocol), the 48 h immobilization of the microcrustacean Daphnia magna (OECD TG 202 performed with the commercially available text kit Daphtoxkit F™ magna, Creasel, Belgium) and the phytotoxicity

NANOBIO&MED 2013

Dendrimers are a class of thoroughly branched polymers characterized by a high chemical versatility. Dendrimers have unique properties such as uniform size, well-defined molecular weight and tunable surface functionality and solubility. Also, the presence of rather large internal cavities makes them interesting for many biological and medical applications [1, 2]. The cytotoxic properties of dendrimers including the influence of surface modification have been the subject of recent scientific publications [3, 4]. Most of the available data found connections between cytotoxicity surface charge and dendrimer generation, which determine cell permeability. In general, cationic PAMAM dendrimers have been associated with higher cytotoxicity than anionic or neutral dendrimers. Also, growing dendrimer generation has been associated with increased toxicity [5]. After therapeutic use dendrimers are excreted. For example, Nigavekar et al. reported that neutral PAMAM G5 dendrimer-based organic nanoparticles resulted in a higher urinary excretion (and lower via faeces) than those bearing positive charge [6]. Once released, the dendrimers would undergo oxidation and photolytic transformation processes, either in water treatment plants or under the action of natural processes. This may lead to the production of secondary pollutants which occurrence and toxicity also needs to be addressed. The goal of this work is to identify and to assess the toxicity of the transformation products of G3-PAMAM-(NH2)32 dendrimer during irradiation and oxidative degradation. Oxidation and irradiation procedures. PAMAM dendrimer G3-PAMAM-(NH2)32 was purchased from Dendritech. The solutions were prepared in pure water (Milipore Mili-Q) with a resistivity of at least 18 Mcm-1 at 25 ºC. Ozone was produced by

roberto.rosal@uah.es

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Advanced Study Institute of Madrid, IMDEA-Agua, Parque Científico Tecnológico, Madrid, Spain 2 Department of Chemical Engineering, University of Alcalá, Alcalá de Henares, Madrid, Spain 3 Department of Environment, Spanish National Institute for Agricultural and Food Research and Technology (INIA), Madrid, Spain

based on seed germination tests following US EPA procedures on Licopersicon esculentum, Lactuca sativa and Lolium perenne [8]. The assessment of reactive oxygen species (ROS) generation was performed by loading cultures of control and treated cells with the fluorescents dyes 2,7dichlorofluorescein diacetate (H2DCFDA) and C4BODIPY, the first serving as indicator for hydrogen peroxide and other ROS, such as hydroxyl and peroxyl radicals and the later being used for evaluating lipid peroxidation. Dichlorofluorescein (DCF), chlorophyll and C4-BODIPY fluorescence was visualised using confocal fluorescence microscopy (Leica TCS SP5). The analytical method developed with LC-ESIQTOF-MS has allowed the identification of the dendrimer G3-PAMAM-(NH2)32 and the structural elucidation of the transformation products generated in the oxidation and irradiation processes. The multiple-charging phenomenon of G3-PAMAM-(NH2)32, associated with ESI, makes it possible the formation of multiply charged ions and provided key advantages in dendrimer identification by assignation of charge state through high resolution of 13C/12C isotopic clusters as well as a mass accuracy below 3.8 ppm for the diagnostic ions. The structures of relevant TP have been elucidated based on mass accuracy, in both full scan and MS/MS modes via IDA, and elemental composition assignation. Fig 1 (left) shows the LCESI-QTOF-MS mass spectrum in full scan mode for G3-PAMAM-(NH2)32. G3 was characterized by a charge state distribution up to +10, 13C/12C isotopic resolution (peak-spacing and isotopic distribution). This figure also presents an overlay of the TIC (total ion chromatograms) with the gradual formation of two relevant transformation products corresponding to the treatment with O3 after 130 min.

[4] Ulaszewska MM, Hernando MD, Uclés A, Rosal

[5] [6]

[7]

[8]

R, Rodríguez A, García-Calvo E, Fernández-Alba AR. Chemical and ecotoxicological assessment of dendrimers in the aquatic environment, in Marinella Farré y Damià Barceló (Eds.) Analysis and Risk of Nanomaterials in Environmental and Food Samples, Comprehensive Analytical Chemistry Volume 59, Chapter 6, 197-233, Elsevier 2012. Naha PC, Davoren M, Casey A, Byrne HJ. Environ. Sci. Technol. 43 (2009) 6864–6869. Nigavekar SS, Sung LY, Llanes M, El-Jawahri A,. Lawrence TS, Becker CW, Balogh L, Khan MK. Pharm. Res., 21 (2004) 476-483. Rosal R, Rodríguez A, Gonzalo MS, GarcíaCalvo E. Appl. Catal. B: Environ. 84 (2008) 48– 57. US EPA. Ecological Effects Test Guidelines. OPPTS 850.4200; Seed Germination/Root Elongation toxicity Test. US Environmental Protection Agency, Washington, DC. EPA712C-96-154 (1996).

Figures

Figure 1: LEFT: LC-ESI-QTOF-MS mass spectrum in full scan mode for G3-PAMAM-(NH2)32 and TIC chromatograms showing the formation of oxidation products. RIGHT: Confocal microscopy images of P. subcapitata exposed to 0.5 mg/L G3NH2 PAMAM during 72 h showing intracellular green BODIPY fluorescence as a result lipid peroxidation. Red fluorescence: chlorophyll autofluorescence. Bottom: overlay image of chlorophyll red autofluorescence and BODIPY green fluorescence and bright field image of the same preparation.

References [1] Suárez IJ, Rosal R, Rodríguez A, Uclés A,

358

Fernández-Alba AR, Hernando MD, GarcíaCalvo E. TrAC Trend. Anal. Chem., 30 (2011) 492-506. [2] Majoros IJ, Baker JR. (2008) Dendrimer-Based Nanomedicine, Pan Stanford Publishing, Singapore, 2008. [3] Menjoge AR, Kannan RM, Tomalia DA. Drug Discov. Today 15 (2010) 171–185.

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Oral Versatile Bionanostructured Materials via Direct Reaction of Functionalized Catechols

Daniel Ruiz-Molina 1 J. Saiz-Poseu 2 B. García J. Sedó1 3 J. Hernando and 3 F. Busqué

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diameter in water, which easily stick to polyester fibers affording stable NP coatings. Even though synthetic polydopamine nanoparticles resembling eumelanin particles that constitute sepia ink have already been described in the literature, to our knowledge this is the first example where chemically-related NPs have been reported to exhibit adhesive properties. On the other side, when this material is dissolved in non-polar solvents such as hexane, robust coatings on a representative variety of substrates, both at the nano-/macroscale are obtained, by means of a quick and ex situ approach without any pretreatment or modification. Whereas catechol monomers bearing a long alkyl chain afford coatings with a persistent hydrophobic character, it was shown that this methodology can be extended to several other catechols with different ring pendant groups, providing varied surface functionalities such as oleophobic/hydrophilic, anti-fouling, anti-bacterial activities and water remediation. Conclusion A new methodology for the fabrication of bionspired catechol-based nanoparticles and coatings is reported. The chemical versatility of the approach allows for a broad range of functionalities that can be added, opening new paths in the realization functional and bioactive materials, an ever growing field of interest. Acknowledgments: This work was supported by MICINN through projects MAT2012-38318-C03-02, MAT2012-38319-C02-01, CTQ2010-15380 and CTQ2009-07469. We are also grateful to Generalitat de Catalunya for project 2010VALOR00039.

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Mussel-adhesive proteins have been the subject of intensive scientific research associated to their remarkable ability to strongly adhere to virtually all surfaces. Albeit diverse in structure, this behavior has been attributed to their varying amounts of the non-essential catecholic aminoacid DOPA. Since this discovery, an ever-increasing number of bioinspired catechol-based polymers have been used for the fabrication of water-resistant adhesives, protective layers, primers for functional adlayers and nanoscale coatings, among others.1 Herein we report a new approach for the preparation of catechol-based materials based on a simple polymerization process in the presence of ammonia,2 in a way reminiscent of melanization reactions. This strategy represents a significant advance in combining many advantages of methods reported previously: ease of preparation, solubility in appropriate solvents and a high ratio of adhesive (catecholic)-to-functional moieties. Experimental methods In a typical experiment, a large molar excess of aqueous ammonia was slowly added under vigorous stirring in the presence of air to a solution of the corresponding monomer (0.2 % , w/v) in methanol, at 40 ºC. Thin-layer chromatography (TLC) was used to follow the reaction, showing that the majority of monomer had already reacted after 3 hours, and consumed quantitatively within 24 hours, after which a dark-brown amorphous solid could be extracted with chloroform and isolated by evaporation. Results and discussion As a proof of concept, the first molecule of choice was a catechol bearing a long alkyl chain. The material resulting from its reaction with ammonia is shown to spontaneously structure in the form of nanoparticles a few hundred nanometers in

druiz@cin2.es

NANOBIO&MED 2013

Centro de Investigación en Nanociencia y Nanotecnología, Campus UAB, 08193, Cerdanyola del Valles,Spain 2 Fundación Privada ASCAMM, Unidad de Nanotecnología (NANOMM), ParcTecnològic del Vallès, Av. UniversitatAutònoma, 23 - 08290 Cerdanyola del Vallès, Spain 3 Chemistry Department, Universitat Autònoma de Barcelona, Campus UAB 08193, Cerdanyola del Vallès, Spain

References [1] Ruiz-Molina et al. et col. Adv. Mat. 2013 DOI: 10.1002/ adma. 201202343. [2] Ruiz-Molina et al. Adv. Mat. 2013 In press. [3] Ruiz-Molina et al. Adv. Funct. Mat. Submitted.

Figures

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Figure 1: Left: (a) Molecular structure of the building block. (b) TEM and (c) SEM images of the polymerization product. (d) Fluorescence emission spectrum of the polymer in aqueous solution (位exc=355 nm, 位det>400 nm) and (inset) fluorescence microscopy image of 2NPs deposited onto glass (位exc=540-552 nm). Right: (a) Schematic representation of the process carried out for coating the substrates with compound 2. (b) TEM images of MWCNT coated with the polymer after being dispersed in a 0.5% (w/v) n-hexane solution for 30 minutes. The green arrows mark the MWCNT wall; the orange arrows point at the coating thickness. (c) Different behavior of blank and treated MWCNT dispersed in water and ethyl acetate.

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Oral Recent therapeutic advances using polystyrene microspheres as delivery systems

Rosario M. Sánchez-Martín

rmsanchez@ugr.es rosario.sanchez@genyo.es

University of Granada, Deparment of Medicinal and Organic Chemistry, Faculty of Pharmacy, Campus Cartuja, s/n, 18071, Granada, Spain GENYO. Centre for Genomics and Oncological Research: Pfizer / University of Granada / Andalusian Regional Government, Av. de la Ilustración, 114 – PTS, 18016, Granada, Spain

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References

N.; How, S.E.; Mittoo, S.; Bradley, M., ChemBioChem, 6 (2005) 1341-1345. [2] Unciti-Broceta, A.; Johansson, E. M.V.; Yusop, M.; Sánchez-Martín, R. M.; Bradley. M., Nature Protocols, 7 (2012) 1207–1218. [3] Sánchez-Martín, R. M., * Alexander, L.; Cardenas-Maestre, J.M.; Bradley, M., Strategies for Microsphere-Mediated Cellular Delivery, Book: Ideas in Chemistry and Molecular Sciences: Where Chemistry meets Life, Wiley-VCH Ed. Pignataro, B. (2010) 117– 140. [4] Sanchez-Martin, R.M.; Cuttle, M.; Mittoo, S.; Bradley, M. Angew. Chem. Int. Ed., 45 (2006) 5472 - 5474. [5] Sanchez-Martin, R. M.*; Alexander, L.; Muzerelle, M.; Cardenas-Maestre, J. M. ; Tsakiridis, A.; Brickman, J. M.; Bradley, M. ChemBioChem, 10 (2009) 1453-1456. [6] Tsakiridis, A.; Alexander, L.; Gennet, N.; Sanchez-Martin, R. M.; Livigni, A.; Li, M.; Bradley, M.; Brickman, J. M., Biomaterials, 30 (2009) 5853-5861. [7] Alexander, L.; Sanchez-Martin, R. M.; Bradley, M. Bioconjugate Chem, 20 (2009) 422-426. [8] Borger, J.G.; Cardenas-Maestre, J.M.; Zamoyska, R.; Sanchez-Martin, R.M.* Bioconjugate Chem,22 (2011) 1904-1908. [9] Cardenas-Maestre, J.M.; Panadero-Fajardo, S.; Perez-Lopez, A.M.; Sanchez-Martin, R.M.* J. Mater. Chem., 21 (2011) 12735-12743. [10] Unciti-Broceta, A.; Díaz-Mochón, J. J.; SánchezMartín, R. M.* and Bradley, M.* Accounts Chem. Res., 45(7) (2012) 1140-1152. [11] Dhaliwal, K.; Alexander, L.; Escher, G.; UncitiBroceta, A.; Jansena, M.; Mcdonalda, N.;

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[1] Sanchez-Martin, R.M.; Muzerelle, M.; Chitkul,

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The search for efficient and safe carrier systems to achieve better availability of therapeutic cargoes at the target site has been a challenging yet exciting area of research. Nowadays many research teams are focused in the development of approaches based in the use of nanodevices as tool to achieve novel therapeutic strategies. We have developed a simple technique for the synthesis of a range of robust and biocompatible functionalized polymer microspheres of highly defined size (100 nm – 500 nm) and loading (Figure 1). [1-2] A range of bioactive cargoes have been conjugated to these particles following a multifunctional strategy and they have been effectively taken up by different type of cells included primary cells and embryonic, neuronal and mesenchymal stem cells (Figure 2).[3] We have demonstrated that these microspheres can efficiently deliver various small molecules (e.g. biosensors) and macromolecules (e.g. proteins).[45] We also show that microsphere internalization does not affect ES cell pluripotency as beadtreated ES cells could be used to generate high contribution chimeric embryos.[6] We have developed several chemical strategies to use these devices as transfection agents (delivery of oligonucleotides such as pDNA and siRNA).[7-10] These devices have been for in vivo cell tracking.[11] Recently we have successfully developed micro heterogeneous catalysts by immobilising palladium nanoparticles on these microspheres and employed them to do chemistry in living systems.[12] These microspheres are inherently attractive as a delivery system due to their lack of toxicity and highly controllable cellular loading.

Cardenas-Maestre, J.M.; Sanchez-Martin, R.; Simpson, J.; Hasletta, C.; Bradley, M. Faraday Discuss., 149 (2011) 107-114. [12] Yusop, M.; Unciti-Broceta, A.; Johansson, E. M.V.; SĂĄnchez-MartĂn, R. M.; Bradley. M. Nature Chemistry, 3 (2011) 239-243. Figures

Figure 1: SEM images of cross linked microspheres.

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Figure 2: Microspheremediated delivery in differentiating Embryonic Stem (ES) cells. Confocal microscopy images of day 7 EBs derived from aEGFP cells.

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Oral Stimuli-Responsive DNA- Nanovalves for Controlled Delivery and Nanodevices

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stimulus such as temperature or pH. These interfaces were incorporated both into mesoporous structures as well as nanoporous membranes [1,2] and upon contacting with the target molecule, they were supposed to cause a change in membrane permeability owing to the significant conformational change of the aptamer receptor molecules. This work will highlight both the capacity of DNA-aptamers for triggering permeability or controlled release, as well as the characterization techniques employed for elucidating and verifying their function and dimensions of the respective conformational changes [3]. Methods: The surface of nanoporous alumina membranes and mesoporous particles was adequately modified for the immobilization of an ATP-aptamer (receptor) as well its unselective mutated form as a negative control. Flouresceine was then used as a tracer molecule in an adjacent feed solution and its permeation across the modified nanoporous membranes monitored as a function of the presence of ATP (target) in the feed. For characterizing the ATP-aptamer responsive membranes, characterization techniques such as quartz crystal microbalance with dissipation monitoring (QCM-D), surface Plasmon resonance (SPR) and dual polarization interferometry (DPI) were employed. The latter proved particularly useful in case where changes were only in the sub-nm range. With regard to the receptor molecules, we opted for an ATP-binding aptamer because ATP with a molar mass of 507,18 g•mol−1 may still be considered a small molecule, and as such posed a challenge to serve as a trigger for conformational changes that would be sufficient to reversibly change the permeability of the overall membrane barrier.

NANOBIO&MED 2013

Approaches to creating stimulus-responsive membranes have been explored for decades for liquid separations or controlled release applications yielding materials whose permeability varies, triggered by a change of pH, temperature, or ionic strength of the adjacent liquid, or the exposure to light, an electrical or a magnetic field. It remains a challenge, however, to mimic the specific and locally acting molecular recognition mechanism found in Nature for triggering a change of permeability in cell membranes, where a specific target-receptor interaction triggers a conformational change of a membrane transporter protein resulting in turn in a variation of the effective diameter of the cell membrane pores. While the incorporation of such transporter proteins is one route to creating artificial molecular stimuliresponsive membranes, a possibly more robust and simpler one is the surface modification of a porous membrane structure with simpler molecules but likewise capable of recognition of a molecular stimulus, such as DNA-aptamers. The capacity of DNAaptamers to reversibly and specifically bind to target molecules of virtually any molecular size while undergoing a conformational change is being explored since very recently for the surface modification of nanoparticless or biosensors1. The challenge for membrane barriers, however, has remained in the application of their molecular recognition principle on a larger substrate area, and most of all to achieve reversibility in its conformational changes for repeated applications. We here report on the performance of selfassembled stimuli-responsive interfaces based on DNAaptamers which respond upon a molecular recognition of a relatively small molecule, adenosinetriphosphate (ATP) rather than a bulk

thomas.schafer@ehu.es

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POLYMAT, University of the Basque Country, Av. Tolosa 72, Donostia-San Sebastián, Spain

Thomas Scháfer V.C. Özalp A. Pinto F.J. Hernadez L. Hernandez and A. Pinto

Results: The self-assembled stimuli-responsive membrane barrier, with its concept depicted in Figure 1, as well as mesoporous particles based on aptamers were capable of reversibly changing their permeability or controlled release upon the molecular recognition of ATP, rather than respond to a bulk stimulus. Negative controls using both mutated aptamers or GTP as a target rather than ATP did not show any significant response of the membrane, whatsoever, proving that the concept proposed did actually rely on a highly selective molecular recognition mechanism as is hitherto mainly known from biological membranes. When the system proposed was furthermore tested under varying medium compositions, such as the presence of serum in order to simulate physiological conditions, no deterioration of the stimuli-responsiveness was observed. The proofof-concept of our membrane design is particularly promising as aptamers as acting receptor molecules can be selected toward virtually any kind of target, ranging from small molecules to proteins and even cells. We will show how we have also successfully applied this modular approach for nanoparticles targeting cancer cells [4], proving that the approach taken addresses a wide range of possible applications in biomedicine or bioseparations in general.

Figures

Figure 1: Schematic of the gate-keeper membranes developed. In absence of the target (ATP), fluoresceine freely permeates the nanoporous membrane; in presence of the target, ATP, the NAaptamer undergoes a drastic conformational change causing a relatice blocking of the pore.

References [1] Özalp, V.C., and Schäfer , T. Chem. Eur. J., 17(36), 9893–9896, 2011. [2] Schäfer, T., Özalp, V. C. in: Responsive

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Membranes and Materials, Eds. D. Bhattacharyya, R. Ranil Wickramasinghe, Sylvia Daunert, Thomas Schäfer, John Wiley & Sons, 2012. [3] Serrano-Santos, M.B., Llobet, E., Özalp, V.C and Schäfer, T. Chem. Commun., 2012, 48, 10087 - 10089 (cover story). [4] Hernandez, F.J., Hernandez, L.I., Pinto, A., Schäfer, T. and Özalp, V.C., Chem. Commun., 2013,49, 1285-1287.

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Invited SERS Microscopy: Tissue Diagnostics with Rationally Designed Nanoparticle Probes

Sebastian Schlücker M. Salehi and M. Schütz

sebastian.schluecker@uni-due.de University of Duisburg-Essen, Physical Chemistry, D-45141 Essen, Germany

References

[2] S. Schlücker, B. Küstner, A. Punge, A. Marx, P.

[3] [4]

[5]

[6] [7]

Ströbel, J. Raman Spectrosc. 37 (2006) 719721. Y. Wang, S. Schlücker, Analyst. 138 (2013) 2224-2238. C. Jehn, B. Küstner, P. Adam, A. Marx, P. Ströbel, C. Schmuck, S. Schlücker, PCCP 11 (2009) 7499-7504. B. Küstner, M. Gellner, M. Schütz, F. Schöppler, A. Marx, P. Ströbel, P. Adam, C. Schmuck, S. Schlücker, Angew. Chem. Int. Ed. 48 (2009) 1950-1953. M. Schütz, B. Küstner, M. Bauer, C. Schmuck, S. Schlücker, Small 6 (2010) 733-737. M. Salehi, D. Steinigeweg, P. Ströbel, A. Marx, J. Packeisen, S. Schlücker, J. Biophoton. (2013), DOI: 10.1002/jbio.201200148.

Figures

NANOBIO&MED 2013

Surface-enhanced Raman scattering microscopy (µSERS) [1] employs target-specific colloidal SERS probes in combination with Raman microspectroscopy. For example, SERS-labeled antibodies allow the selective and sensitive localization of the corresponding antigen in tissue specimens [2]. The physical and chemical properties of the colloidal SERS probes are crucial for the success of SERS microscopic experiments [3]. Stability and robustness, sensitivity as well as steric accessibility for bioconjugation are few very important aspects. Figure 1 left shows a hydrophilic SERS probe stabilized by many short and few longer hydrophilic spacer units [4]. A second approach, based on the glass encapsulation for stabilizing SERS labels, is schematically depicted in Figure 1 right. This type of SERS probes was optimized for red laser excitation in order to improve image contrast by minimizing unwanted autofluorescence of biological specimens [5]. Two different routes to the glass encapsulation of a complete monolayer of Raman reporters will be described [5-6]. Small clusters (dimers, trimers) of gold nanocrystals even exhibit single-particle sensitivity and enable rapid mapping experiments with only 30 msec acquisition time per pixel [7]. Future directions of this innovative Raman/SERS microspectrosopic technique for tissue-based tumor diagnostics will be discussed.

Figure 1: (Left) SERS labels [3] stabilized by many short and few longer hydrophilic spacer molecules for controlled bioconjugation [4]. (Right) SERS nanoprobes optimized for red laser excitation for minimizing tissue autofluorescence. Gold/silver nanoshells are covered by a complete self-assembled monolayer (SAM) of Raman labels. The SERS particle is protected and stabilized by a glass shell [5-6].

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[1] S. Schlücker, ChemPhysChem 10 (2009) 13441354.

Oral Current research strategies for toxicity assessment of engineered nanomaterials. Lessons learned from European Projects

Blanca Suárez-Merino F. Goñi de Cerio C. Aristimuño and P. Heredia

suarezb@gaiker.es GAIKER Technology Center, Parque Tecnológico de Zamudio Ed. 202, 48170, Zamudio, Vizcaya, Spain

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The rate at which novel nanomaterials are entering the marketplace and their unlimited economic potential is not being followed by updated regulation regarding the use of nanomaterials and their implications for human and environmental safety. The consequences of this gap of knowledge are at least two fold, on one hand consumer´s confidence is at stake, on the other there is a risk to lose an opportunity for innovation. At present there are a large number of uncertainties regarding the use of traditional toxicology to evaluate the toxicity of nanomaterials, which, due to their special physical characteristics, require further assessment when considering the particular methodology to be used. Furthermore, these new properties may be able to alter the absorption and transport capacity of nanoparticles across membranes. There is also a potential for nanoparticles to accumulate in organs, enter into blood circulation or even cross biological barriers. In this context GAIKER-IK4 is involved in the development of novel methodologies to perform in vitro toxicological evaluation combining technologies such as flow cytometry, confocal microscopy and nanogenotoxicity to produce an in vitro tool-kit to accurately assess nanotoxicity. Our experience is based on our participation and leading roles in International Projects dealing with safety and efficacy assessment of nanomaterials from a Pharma, Cosmetic and Occupational Health point of view [1,2]. Our strategy aims at the core of engineered nanomaterial development, assisting in nanomaterial design by providing a fast and reliable evaluation of nanoparticle toxicity in the same fashion as any other conventional drugs are assessed for toxicity at their preclinical stage.

References [1] Serri R, Iorizzo M, Cosmeceuticals: focus on

topical retinoids in photoaging. Clin Dermatol 26 (6) (2008) 633-635. [2] Halevy S Dead Sea bath salt for the treatment of psoriasis vulgaris: a double-blind controlled study. Journal of the European Academy of Dermatology and Venereology 9. (1997) 237242.

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Oral

College of Engineering, Swansea University, SA2 8PP Swansea, UK College of medicine, Swansea University, SA2 8PP Swansea, UK

Human chorionic gonadotropin (hCG), a 37 kDa glycoprotein hormone [1], is an important biomarker for pregnancy and one of the most important tumor markers for several cancers [2]. Most modern assay systems for hCG are optical based ELISAs. Though advances have been made in miniaturization of sensor diagnostics, in point of care detection, and even automated/continuous monitoring systems, the desired level of sensitivity for early detection and monitoring for cancer has not yet been achieved. A novel chemically-modified graphene diagnostic sensor has been developed for ultrasensitive detection of a hCG (human chorionic gonadotropin) biomarker. Multi-layer epitaxial graphene (MEG), grown on silicon carbide substrates, has been patterned using electron beam lithography to produce channel based devices. Silicon Carbide (SiC) is a suitable substrate for graphene growth [3-7]. Graphene is grown by sublimating the silicon from the SiC substrate. Graphene, essentially a single atomic layer of graphite, has some exceptional electronic properties such as carrier mobilities of more than 15000 cm2 V-1s-1 at room temperature and ballistic transport of carriers [4]. Modifying the growth conditions yields variations in the graphene layer thickness and consequently its electronic properties. Diagnostics based on bio-functionalised semiconductor devices are an important development in ultrasensitive sensors for early detection and monitoring of disease biomarkers. The chemical sensor uses chemically-modified graphene channels, functionalized with a “bioreceptor” antibody to detect the presence of the disease target biomarker, anti-hCG. It is then able to interact with the surface-bound bioreceptor and produce a change in the charge properties of the

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r.s.r.teixeira.607010@swansea.ac.uk

biosensor device. This surface charge modification is measured by amperometry on our graphenebased channel devices. The attachment of the anti-hCG onto the graphene surface requires two parallel processes: the surface modification and the antibody modification. The surface modification is performed by firstly terminating the graphene with -OH groups using the Fenton reaction [8], then reacted with 3Aminopropyl-triethoxysilane (APTES) [9] in order to obtain an amine-terminated surface. This is illustrated in Figure 1 (a to d) where the presence of the silane and the carbon chain from the Aptes molecule are clearly detected using XPS. Amine groups, along with oxygen in the C-O-Si configuration were also detected (Table 1). In order to be able to react with the surface amine groups, the carboxylic acids on the antibody are activated. However, in order to prevent the antibody from cross-linking, the majority of amine groups on the antibody are blocked using Di-tertbutyl dicarbonate. The antibody can now be reacted with the amine on the surface. The groups blocking the amines on the antibody are subsequently removed by mild acidic treatment. The sensitivity of the obtained device was then tested against varying concentration of hCG by measuring the resistance across the modified graphene channel (Figure 2). The obtained result showed a sensitivity of the sensor of 142 Ω/(ng/ml), and a detection limit around 1ng/ml.

References [1] J. Chen, J.H. Tang, et al. Biomaterials 27 (2006) 2313–2321.

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Sofia Teixeira 1 A. Castaing 1 G. Burwell R.S. Conlan2 and 1 O.J. Guy

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Graphene Biosensor for Detection of hCG Biomarker

[2] M. Aizawa, A. Morioka, et al. Anal. Biochem. 94 (1979) 22–28. [3] O.J. Guy, M. Lodzinski, et al. Appl. Surf. Sci.,

Vol. 254, pp.8098-8105, 2008. [4] C. Berger, Z. Song, et al. Science, Vol. 312, pp.1191-1196, 2006. [5] A. Charrier, A. Coati, et al. J. Appl. Phys., Vol. 92,pp.2479-2484, 2002. [6] C. Berger, Z. Song, et al. J. Phys. Chem. B, Vol. 108, pp.19912-19916,2004. [7] A. Castaing, O.J. Guy, et al. Mater. Sci. Forum, Vols. 615-617, pp.223-226, 2009. [8] R.H. Bradley, K. Cassity, et al. Applied Surface Science 258 (2012) 4835–4843. [9] J. Kathi, K. Y. Rhee. J Mater Sci (2008) 43:33– 37.

Figure 1 (d): XPS core level spectrum of the C1s peak of a graphene sample after functionalisation (a) measured data, (b) fitted graphene peak(c) fitted SiC peak, (d), fitted APTES peak..

Figures

Figure 2: hCG concentration plotted as a function of channel resistance across a 100 μmx 4mm graphene channel.

Figure 1 (a): XPS core level spectrum of the Si2p peak of an epitaxial graphene sample (grown on SiC) before functionalisation (a) measured data, (b) fitted peak attributed to SiC.

Tabla: Description of bonds and atomic abundances calculated from the fitted components of the C, Si and N core peaks from XPS measurement before and after functionalisation with APTES.

Figure 1 (b): XPS core level spectrum of the Si2p peak of an epitaxial graphene sample after functionalisation (a) measured data, (b) fitted peak attributed to SiC and (c) fitted peak attributed to the silicon atom of the APTES molecule

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Figure 1 (c): XPS core level spectrum of the C1s peak of a graphene sample before functionalisation (a) measured data, (b) fitted SiC peak and (c) fitted epitaxial graphene peak.

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Oral

Nano clays as sepiolite and clinoptilolite, have attracted increasing interest in biomedicine for their potential applications as nanovehicles. However, no attempts have been made to evaluate the potential undesirable effects of sepiolite and clinoptilolite nanoparticles. The cytotoxicity of this two nano clays with different chemical structures was systematically evaluated in various macrophages types, such as human peripheral blood, mice bone marrow and cell line 264,7, by measuring cell viability and apoptotic pathway of death. In vivo evaluation was performed exposing CD1mice to suspensions of nano sepiolite and clinoptilolite at different concentrations (1, 2.5 and 5 mg/Kg of weight) by different ways (cutaneous, subcutaneous, intramuscular and intraperitoneal). No significant cytotoxic or toxic effects could be seen in both cases (in vitro and in vivo), but nano sepiolite was determined to be slightly more toxic than clinoptilolite in terms of cell viability and inflammation, this may be due to the differences in the structure. It is, therefore, expected that nano clays could be promising candidates for novel inorganic drug delivery carriers. Introduction The origin of nanoscience and nanotechnologies is often attributed to a concept advanced by Nobel Laureate Richard P. Feynman, who in a 1959 lecture at the California Institute of Technology, stated: “There is plenty ofroom at the bottom. Many of the cells are very tiny, but they are active: they manufacture substance; they walk around; they wiggle: and they do all kinds ofmarvelous thingsall on a very small scale. Also they store information. Consider the possibility that we too can make things very small which does what we want when we want-and that we can manufacture an object that maneuvers at that

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yanistoledano@gmail.com

level”. [1] Nanoparticles are used in various fields such as photonics, catalysis, magnetics, and biotechnology including cosmetics, pharmaceutics, and medicines. In particular, research interest has focused on their potential as drug delivery nano vehicles and nano medicines [2,3] It is, therefore, needed to answer the questions on their safety issue by performing vigorous toxicological evaluation based on various models [4-7], and to understand their interaction mechanism as well. Such a toxicological study can provide not only the critical information on the biological applications of nanoparticles, but also help to avoid any undesirable effects [8]. However, only a few studies have evaluated the safety of nano-sized materials and their potential adverse effects on biological systems [9]. Materials and methods. Parasite culture Entamoeba histolytica HM1-IMSS trophozoites were axenically grown in TYI-S33 medium supplemented with each nano clay so that the final concentrations were as follows: 1, 10, 50, and 100 mg/ml. Amoebic trophozoites viability was assessed employing two different methods, (1) vital marker trypan blue and (2) carboxyfluorescein diacetate (CFDA Vibrant kit) and propidium iodide done at 24, 28 and 72 hours under a microscope using a haemocytometer. Cellular assays Macrophages from human peripheral blood, for each experiment 1x105 cells per well were placed in 96 well-plate with 100μl of RPMI 1640 supplemented and enough nanoclay to reach 0.1, 1, 10 and 100mg/ml of medium in each well. Apoptosis and necrosis were analysed each 12 hours for 48 hours by flow cytometry. In vivo tests CD1 mice were treated orally, cutaneously, subcutaneously, intramuscular and intraperitoneal with 1, 2.5 and 5mg of nano clay/Kg of weight and

NANOBIO&MED 2013

Universidad Nacional Autónoma de México, Circuito exterior s/n, Ciudad Universitaria, CP 04510 México D.F., México.

Yanis Toledano-Magaña L. Flores-Santos G. Montes de Oca-Ramírez A. González-Montiel and J. C. Carrero

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Biocompatibility of nanoclays for future applications in biomedicine

analytes were measured for kidney and kiver function. Results There is no significant effect in the cultures of Entamoeba histolytica treated with clinopiloite but the one that can be seen is dose and time dependent like for sepiolite but for this one the effect is significant for the higher concentrations (50 and 100mg/ml) it can be associated with the structure of the clays cause it is known that sepiolite is a fibrous clay until clinoptilolite have octahedric structure (Figure1). In the case of HPB (human peripheral blood) macrophages the effect is significant for the two clays at the higher concentrations (10, 100μg/ml) but is largest the percentage of cells affected with nano sepiolite (25%) than with nano clinoptilolite (20%) (Figure2). For in vivo tests we observed the accumulation of nano clays without inflammation (Figure3), but orally the mice stomachs treated with nano sepiolite have a size reduction and black color in some areas (Figure4) however kidney and liver function are normal (Figure 5). Conclusions Nano clinoptilolite seems to be a ciocomativle nano clay that in the case of nano sepiolite more studies are necessary but the effect is higher in cultures and animals.

Figures

Figure 1: Left A Cultures of Entamoeba histolytica treated with nano clinoptilolite. B Cultures of Entamoeba histolytica treated with nano sepiolite. Right Analyze with CFDA and PI that shows in green the live cells and in red the nucleus of dead cells.

Figure 2: A Macrophages of HPB treated with nano clinoptilolite. B Cultures of macrophages of HPB treated with nano sepiolite. Cells were analyzed by flow cytometry in a FacsCanto cytometer.

Figure 3: Accumulation of nano clays in the administration areas in CD1 mice.

References [1] Feynman RP, Leighton RB, Sands M. The

[2] [3] [4] [5] [6]

[7] [8]

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[9]

Feynman Lectures on Physics, vol. 1. Reading MA: Addison Wesley Publishers; 1961. S.M. Moghimi, A.C. Hunter, J.C. Murray, Faseb. J. 19 (2005) 311–330. U. Pison, T. Welte, M. Giersig, D.A. Groneberg, Eur. J. Pharmacol. 533 (2006) 341–350. H.M. Kipen, D.L. Laskin, Am. J. Physiol. Lung Cell Mol. Physiol. 289 (2005) L696–697. A. Nel, T. Xia, L. Madler, N. Li, Science 311 (2006) 622–627. G. Oberdorster, E. Oberdorster, J. Oberdorster, Environ. Health Perspect. 113 (2005) 823–839. M.C. Powell, M.S. Kanarek, W. M. J. 105 (2006) 16–20. K. Thomas, P. Sayre, Toxicol. Sci. 87 (2005) 316–321. R.F. Service, Science 304 (2004) 1732–1734.

Figure 4: Effect on the stomach morphology and colouration of CD1 mice treated with nano sepiolite.

Figure 5: Analytes for liver and kidney fuction that are in the normal range.

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Invited Visulaization of proteins in solution in situ

Rupert Tscheliessnig S. Ho Shin and A. Jungbauer

rupert.tscheliessnig@boku.ac.at ACIB, Muthgasse 11, 1190 Vienna, Austria

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References

[2] [3]

[4]

[5]

[6]

Angew. Chem. Int. Ed. 1999, 38, 1034. b) U.B. Sleytr, P. Messner, D. Pum, M. Sara, Crystalline Bacterial Cell Surface Protein, Landes/Academic Press, Austin, 1996. c) J. Lechner, F. Wieland, Annu. Rev. Biochem. 1989, 58, 173-194. S. Mann, Angewandte Chemie-International Edition 47, 5306-5320(2008). a) D. Moll, C. Huber, B. Schlegel, D. Pum, U. B. Sleytr, M. Sara, Proceedings of the National Academy of Sciences of the United States of America, 2002, 99 ,14646. b) B. Aichmayer, M. Mertig, A. Kirchner, O. Paris, P. Fratzl, Adv. Mater. 2006, 18, 915. c) S. S. Mark, M. Bergkvist, X. Yang, L. M. Teixeira, P. Bhatnagar, E. R.Angert, C. A. Batt, Langmuir 2006, 22, 3763. c) W. Shenton, D. Pum, U. B. Sleytr, S. Mann, Nature, 1997, 585. a) J. E. Norville, D. F. Kelly, T. F. Knight, A. M. Belcher, T. Walz, J. Struct. Biol . 2007 , 160 , 313-323. b) H. Engelhard, J. Struct. Biol. 2007, 160 , 115-124. c) H. Engelhard, J. Struct. Biol. 2007 , 160 , 190-199. S. Chung, S.-H. Shin, C. R. Bertozzi, J. J. DeYoreo, Proceedings of the National Academy of Sciences of the United States of America, 2010 . 107 (38) 16536-16541. a) C. Horejs, D. Pum, U. B. Sleytr, R. Tscheliessnig, J. Chem. Phys. 2008, 128 , 065106. b) C. Horejs, H. Gollner, Di. Pum, U. B. Sleytr, H. Peterlik, Al. Jungbauer, R. Tscheliessnig, ACS Nano, 2011, 5 (3), pp 2288â&#x20AC;&#x201C;2297. c) C. Horejs, D. Pum, U. B. Sleytr, H. Peterlik, A. Jungbauer, R. Tscheliessnig, J. Chem. Phys. 2010, 133 , 175102.

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[1] a) U.B. Sleytr, P. Messner, D. Pum, M. Sara,

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Time dependent visualization of proteins in situ, e.g. their degradation during manufacturing and storage, the impact of salts upon their conformations are of broad interest in industrial biotechnology. The interest gets broader and arouses interest wihin the field of material sciences if it comes to self assemblies. The outmost of cells of a broad spectrum of bacteria and archae are covered by layer proteins (S-layers). Consequently these are strongly involved in metabolic processes in response to the environment, serving numerous functions including structural stabilization, cell adhesion, pathogenicity, drug resistance and mineralization [1]. These two-dimensional crystals of S-layers have been recognized as a distinct class among numerous protein architectures found in nature [2, 4] that present various lattice periodicities commensurate with the dimensions of synthetic nano-materials, such as quantum dot and carbon nano-tubes. These long-range order self assemblies attractive bio-molecules for nano-scale templates for prospective nano-material engineering [3]. The assembly process in particular its detailed mechanism is understood if at all, poorly. We combine Differential Scanning Calorimetry, in situ atomic force microscopy [5], Small Angle X-ray Scattering [6], Transmission electron microscopy as well as tomography with reversed Molecular dynamic simulations and reverse Monte Carlo simulations, to model possible crystallization pathways and conformational changes in situ.

Figures

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Figure 1: Gray data refer to 3D experimental data of SbpA unit cell. a-c) Three complementary conformations of an SbpA monomer are superimposed. Pink circles refer to possible positions of secondary structure elements.

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Oral Magnetoresponsive hybrid microgels: close packed magnetite nanoparticles core - double layer polymeric shells with anionic or cationic functionalities 1

Rodica Turcu 1 I. Craciunescu 2 V. Socoliuc A. Petran1 2 C. Daia and 2 L. Vekas

rodica.turcu@itim-cj.ro

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microgels with cation exchange (CEX) or anion exchange (AEX) functionalities have been obtained by coating the NPCs with two polymer shells using layer by layer free radical polymerization of either N-isopropylacrylamide and acrylic acid (M-NIPAAAc) or N-isopropylacrylamide and 3acrylamidopropyl)-trimethylammonium chloride (M-NIPA-APTAC) in the presence of the crosslinker N,N-methylene bis-acrylamide and oxidant ammonium persulfate. The TEM image of NPCs coated with SDS, figure 1 shows that magnetite nanoparticles stabilized with oleic acid are densely packed into spherical clusters coated with SDS with sizes in the range 100-500 nm. Dynamic light scattering and zeta potential measurements of the microgels gives values of the mean hydrodynamic diameter, Dh in the range 517-914 nm and zeta potential values between -23mV and -37mV for CEX microgels and between +35 mV and +39 mV for AEX microgels. The magnetization vs. applied magnetic field at room temperature for NPCs stabilized with SDS and for magnetic microgels shows superparamagnetic behavior, the saturation magnetization has relatively high values in the range 46-56 emu/g, figure 2. The chemical surface analysis of magnetic nanoparticle clusters and magnetic microgels was performed by X-ray Photoelectron spectroscopy (XPS). The coating of NPCs stabilized with SDS with double layer of polymers poly(N-isopropyl acrylamide) and polyacrylic acid is evidenced by the characteristic peaks in XPS spectra: (i) amide group N-C=O (286 eV) in C1s spectrum, NH (399 eV) in N1s spectrum, C=O (532.5 eV) in O1s spectrum, which are specific for pNIPA; (ii) carboxyl group O-C=O in C1s and O1s spectrum, specific for pAAc. XPS spectra of AEX magnetic

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Magnetic nanoparticles (MNPs) well separated or in clusters, with proper surface functionalization usually dispersed in liquid carriers or embedded/encapsulated in polymeric networks, are the basic building blocks of a large variety of multifunctional carriers. The encapsulation of magnetic nanoparticles into polymeric micro/nanogel systems gives rise to hybrid magnetic carriers that combine the features of a polymeric gel with the interesting properties of magnetic nanoparticles and make them suitable for applications such as magnetically targeted drug delivery, hyperthermia treatments, magnetic resonance imaging contrast enhancement, magnetic carriers in bioseparation equipments [12]. We report the synthesis and characterization of magnetic microgels based on controlled clustering of hydrophobic magnetite nanoparticles and coating with different polymeric shells of poly(Nisopropylacrylamide), polyacrylic acid, poly(3acrylamidopropyl)-trimethylammonium chloride. The oil in water miniemulsion method [3] was applied for the preparation of magnetic nanoparticles clusters (NPCs) using toluene based magnetic nanofluid (MF) containing Fe3O4 nanoparticles stabilized with a hydrophobic layer of oleic acid. The MF was added to the aqueous solution containing sodium dodecyl sulphate (SDS) as surfactant and treated ultrasonically to obtain small stable droplets with NPCs. The as prepared magnetic miniemulsion was heated to remove the toluene and then was carefully washed several times with methanol-water mixture, magnetically separated and redispersed in water. Stable aqueous dispersions of magnetic nanoparticles clusters coated with the surfactant SDS were used for encapsulation into polymers. Magnetic

NANOBIO&MED 2013

National Institute R&D for Isotopic and Molecular Technologies, 65-103 Donath Street, 400293 Cluj-Napoca, Romania 2 Romanian Academy-Timisoara Branch, Center for Fundamental and Advanced Technical Research, Lab. Magnetic Fluids, Timisoara, Romania

microgel, confirm the double layer polymer coating of SDS stabilized NPCs with poly(Nisopropyl acrylamide) and poly(3-Acryloamidopropyl trimethylammonium chloride), figure 3. The CEX and AEX magnetic microgels exhibit good stability, high saturation magnetization/large magnetic moment, response to moderate magnetic fields, being promising materials for applications in high gradient magnetic separation and nanomedicine. Acknowledgements: Support by the European project FP7 No. 229335 MAGPROÂ˛LIFE and the Romanian project 83EU/2010 is acknowledged.

References

Figure 3: High resolution XPS spectra for C1s, O1s, N1s, S2p, core levels from AEX microgel M-NIPA-APTAC obtained by coating of SDS stabilized NPCs with poly(N-isopropyl acrylamide) and poly(3Acryloamido-propyl trimethylammonium chloride).

[1] . Lu, M. Ballauff, Progress in Polymer Science, 36 (2011) 767-792. [2] N.S. Satarkar, D. Biswal, J.Z. Hilt, Soft Matter, 6

(2010) 2364-2371. [3] P. Qiu, C. Jensen, N. Charity, R. Towner, C.

Mao, J. AM. CHEM. SOC. 132 (2010) 17724â&#x20AC;&#x201C; 17732. Figures

Figure 1: TEM images of NPCs stabilized with SDS prepared by oil in water miniemulsion method.

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Figure 2: Magnetization curves at room temperature for NPCs and magnetic microgels M-NIPA-AAc (CEX), M-NIPA-APTAC (AEX).

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Oral Characterization of Functionalized, Magnetic Nanoparticles

Barbara Unterauer M. Rohn and M. Mühlberger

Profactor GmbH, Im Stadtgut A2, 4407 Steyr-Gleink, Austria

barbara.unterauer@profactor.at

Acknowledgements: This work was supported by Die Österreichische Forschungsförde rungsge sellschaft FFG. Part of this work was supported by the NanoInk and NanoShape projects of the NSI project cluster in the Austrian nanoinitiative (www.nanoscience.at). Part of this work was also supported by the MacroFun project in the COMET initiative (www.macrofun.tugraz.at).

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References [1] Kovalenko M.V., Bodnarchuk M.I., Lechner

R.T., Hesser G., Schäffler F., Heiss W., Journal of American Chemical Society 129 (2007) 6352.

Figure 1: Reaction scheme of the particle synthesis. These particles are characterized with AFM.

NANOBIO&MED 2013

Figures

Figure 2: Size and size distribution of particles on the example of a 1μm AFM image.

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Nanoparticles play an important role in various fields like pharmaceuticals, advanced materials, environmental detection and monitoring. Obviously there is not a single best technique to characterize nanoparticles, but AFM is a very suitable technique offering qualitative and quantitative information on the particle. In this study we are dealing with the characterization of iron-oxide nanoparticles. They were synthesized in a batch process based on the protocol described by Kovalenko et al [1] and shown in Figure 1 below. After particle formation the oleate shell was replaced by hydrophilic ligands. In one special case we functionalized them with a dye to create a so-called laccase sensor. For the characterization we were interested in qualitative and quantitative information on physical properties like size, morphology and magnetic properties and statistical information on size distribution. We will show the particle synthesis, the various functionalization products, the sample preparation for the AFM measurements and the results of the measurements.

Oral Ruben Van Roosbroeck 1 T. Stakenborg 1 W. Van Roy J. Lammertyn2 and 1 L. Lagae

Ruben.Vanroosbroeck@imec.be

Imec, Kapeldreef 75, Leuven, Belgium KUL (Mebios), Kasteelpark Arenberg 30, Leuven, Belgium

Magnetic nanoparticles show great promise and are widely investigated as agents in various biomedical applications including analyte detection, biosensing, drug delivery and imaging. A particular field of interest is magnetic resonance imaging (MRI) where such particles can serve as T2 contrast enhancers. Most of the reported research related to MRI is focussing on superparamagnetic nanoparticles, synthesized by bottom-up wet chemistry methods[1]. These magnetic particles are, however, restricted to the superparamagnetic limit to avoid magnetic remanence and aggregation in the absence of a magnetic field. This restriction therefor implies a size limit (20-25 nm) and consequently a limited magnetic moment per particle. The ideal particle diameter for contrast agents in MRI-experiments unfortunately exceeds this limit[2]. Moreover, bottom-up synthesis often results in polydisperse particle suspensions. To overcome these issues, synthetic antiferromagnetic (SAF) particles are proposed here as alternative T2 contrast agents for MRI applications. These particles are composed of two ferromagnetic layers, separated by a non-magnetic layer (Figure 1). Using the interlayer exchange coupling effect, synthetic antiferromagnetism can be produced which mimics superparamagnetic behavior (i.e. no magnetic moment in the absence of a magnetic field). Size and shape can be controlled through a range of litohgraphic methods to produce highly magnetic and monodisperse particle suspensions. To fabricate the SAF particles, we used colloidal lithography with monodisperse polystyrene beads (60-200 nm) serving as an etch mask. By controlling the size of the polystyrene beads, the final SAF particles could be fine-tuned in size. After synthesis on wafer

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1,2

Synthetic Antiferromagnetic Nanoparticles As Contrast Agents In MRI

scale and a lift-off step, the particles were suspended in aqueous solution and chemically stabilized. As intended, the particles showed no magnetic remanence despite the fact that they are composed of ferromagnetic materials (Figure 2). To show the performance of the SAF particles as contrast agents simulations were performed and experimental relaxivities were determined. Using simulations the effect of size, composition and shape of the particles on the relaxivity was demonstrated. Second, experimental relaxivities r2 up to 350 s-1mM-1 were achieved for NiFe-based particles which is competitive with conventional superparamagnetic nanoparticle suspensions[3]. These proposed SAF particles, behaving like superparamagnetic particles without being restricted in size, show a great potential for MRI applications.

References [1] Hilgner I, Kaiser WA, Nanomedicine 7 (2012) 1443-1459. [2] Vuong QL, Gillis P, Gossuin Y, Journal of

Magnetic Resonance 212 (2011) 193-48. [3] Vuong QL, Berret JF, Fresnais J, Gossuin Y,

Sandre O, Advanced Healthcare Materials 1 (2012) 502-512.

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Figures

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Figure 2: SQUID-measurement of suspended synthetic antiferromagnetic nanoparticles with a mean diameter of 100 nm. The schematics show the magnetization of the magnetic layers inside the particle. At zero field no magnetic remanence is observed due to the SAF-coupling of the magnetic layers.

NANOBIO&MED 2013

Figure 1: Electron microscopic image of the fabricated synthetic antiferromagnetic nanoparticles with a mean diameter of 100nm. The inset (upper right) shows a magnification of a single particle. The SAF particle is composed of two NiFe layers (black) in the center separated by a thin Ru-spacer (bright). A gold layer on top and bottom (thick bright layers) surround the SAF stack.

Oral Ladislau Vékás 1 1 V. Socoliuc , F. Bălănean 1 1 A. Moaca , C. Daia D. Susan-Resiga1, O. Marinică1 1 2 N.C. Popa , T. Borbáth 2 3 T. Boros and R. Turcu

vekas@acad-tim.tm.edu.ro vekas.ladislau@gmail.com

Romanian Academy-Timisoara Branch (RATB), 24 Mihai Viteazul Str., Timisoara, Romania 2 ROSEAL Co., N. Balcescu Str. 5A. Odorheiu Secuiesc, Romania 3 National Institute of R&D for Isotopic and Molecular Technologies (NIIMT), 65-103 Donath Str., Cluj-Napoca, Romania

High magnetic moment nano – or micron sized carriers with superparamagnetic behavior and various added functionalities provide advanced therapeutic, diagnostic or bioprocessing capabilities [1]. The strongly magnetoresponsive character and appropriate design of surface properties are among the main requirements for nano- or micro-adsorbents to be used in magnetic bio-separation processes, such as protein purification [2] involving high gradient magnetic separation (HGMS) step [3]. The building blocks for the fabrication of magnetoresponsive microcarriers are the subdomain magnetite nanoparticles surface coated by biocompatible molecular layers, such as carboxylic (lauric, myristic, oleic) acids, which ensure their stabilization-dispersion in an appropriate carrier liquid, to obtain stable magnetic nanofluids - the primary materials for the envisaged magnetic nanocomposites. In this work we report on kg scale synthesis of magnetite nanoparticles with biocompatible coating used as primary particles for the preparation of magnetoresponsive hybrid nanocomposites designed for magnetic separation [4] or drug targeting [5]. Controlled clusterization of magnetite nanoparticles from light hydrocarbon or water based magnetic nanofluids is a widely applied procedure to synthesize polymer-magnetic nanoparticles hybrid microspheres with sizes well above 50 nm having both high magnetic moment and superparamagnetic behavior [4, 5]. The synthesis procedures of magnetite nanofluids (fig.1) using the chemical co-precipitation method developed on laboratory level [6] were optimized and scaled up to achieve the main characteristics required for the preparation of magnetoresponsive microspheres. The procedures

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Hydrophobic and hydrophilic magnetite nanoparticles: non-polar and polar magnetic nanofluids designed for magnetic carriers manufacturing

were validated by using the data of TEM, DLS, SLS, VSM, magnetogranulometry and rotational rheometry investigations: mean particle size 6-9 nm (figs.2 and 3), biocompatible surface coating (vegetable origin oleic acid; Merck product), high colloidal stability (reduced fraction of small agglomerates; hydrodynamic mean size below 50 nm, figs.4 and 5), well screened magnetic dipoledipole interactions between particles also at close packing (fig.6; ϕ= 0.2 corresponds to approx. 0.5 hydrodynamic volume fraction); superparamagnetic behavior and high specific magnetic moment of surfacted magnetite nanoparticles (fig.7; approx. 50 emu/g), reduced costs. Acknowledgement: Financial support by the European project FP7 No. 229335 MAGPRO²LIFE and the Romanian project 86EU/10CF/2010 is acknowledged.

References [1] L.H. Reddy, J.L. Arias, J. Nicolas, P. Couvreur, Chemical Reviews, 112 (2012) 5818-5878. [2] C. Müller, K. Wagner, K. Frankenfeld, M.

Franzreb, Biotechnology Lett., 33(5) (2011) 929-936. [3] G. N. Brown, C. Müller, E. Theodosiou, M. Franzreb, O.R.T. Thomas, Biotechnology and Bioengineering, DOI: 10.1002/bit.24842 (2013). [4] M.S.A. Darwish, U. Peuker, U. Kunz, T. Turek, J. Mater. Sci., 46 (2011) 2123-2134. [5] L. Vékás, EtelkaTombácz, RodicaTurcu, I. Morjan, M.V. Avdeev, Theodora KrasiaChristoforou, V. Socoliuc, in: Nanomedicine-

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Basic and Clinical Applications in Diagnostics and Therapy (Ed. ChristophAlexiou) Karger Publ. Basel, Switzerland, 2 (2011) 35-52. [6] L. Vekas, M.V. Avdeev, D. Bica, in: NanoScience in Biomedicine (Ed. Donglu Shi, Springer USA, 2009) 645-70. Figures Figure 5: Hydrodynamic size distribution of hydrophobic magnetite NPs (Fe3O4.OA).

Figure 6: Dependence of magnetic mean size and standard deviation on the solid volume fraction of magnetite NPs dispersed in hydrocarbon carrier. Figure 2: TEM of magnetite NPs dispersed in hydrocarbon carrier liquid.

Figure 3: Size distribution of magnetite NPs.

NANOBIO&MED 2013

Figure 1: Magnetic nanoparticle synthesis procedures-main steps.

Figure 7: Full magnetization curve for hydrophobic magnetite organosol (Fe3O4.OA).

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Figure 4: Hydrodynamic size distribution of hydrophilic magnetite NPs (Fe3O4.(OA+OA)).

Invited Precisions synthesis of nanocrystals and their use in biomedical applications

Horst Weller

weller@chemie.uni-hamburg.de Department of Chemistry, University of Hamburg Center for Applied Nanotechnology Hamburg (CAN) Interdisciplinary Nanoscience Center Hamburg (INCH) The Hamburg Center for Ultrafast Imaging (CUI) Grindelallee 117 - 20146 Hamburg, Germany

380

We report on the precision synthesis of fluorescent and magnetic nanocrystals using a preparative flow reactor. Experimental design is used to determine the crucial parameters and their influence on particle growth and size distribution. In the second part of the talk, we will present biological applications of nanocrystals. In particular, we will show a comparison of different shell materials in respect of unspecific cell adhesion and uptake and discuss parameters controlling these processes. Nanocrystals, which were equipped with cross-linked block-copolymer ligand shells or encapsulated by emulsion polymerization show extraordinary low unspecific cell interactions. We present various techniques for bio-conjugation with recognition molecules and show examples for specific cell and tissue targeting. In-vitro and in-vivo fluorescence and MRI data will be discussed

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Invited Biomedical applications of magnetic nanoparticles: from cell imaging to tissue engineering

Claire Wilhelm

claire.wilhelm@univ-paris-diderot.fr

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS and Université Paris Diderot, France

References

[2]

[3]

[4]

[5]

[6]

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3161. Levy M, Luciani N, Alloyeau D, Elgrabli D, Deveaux V, Pechoux C, Chat S, Wang G, Vats N, Gendron F, Factor C, Lotersztajn S, Luciani A, Wilhelm C, Gazeau F, Biomaterials. 32 (2011) 3988. Lesieur S, Gazeau F, Menager C, Wilhelm C. J. Mat. Chem. 21, 14387-14393 (2011). Lartigue L, Wilhelm C, Servais J, Factor C, Dencausse A, Bacri J-C, Luciani N, Gazeau F. ACS Nano, 6 (2012) 2665. Wilhelm C, Phys Rev Lett 101 (2008) 028101. Robert D, Nguyen TH, Gallet F, Wilhelm C, PLoS one 5, (2010) e10046. Robert D, Pamme N, Conjeaud H, Gazeau F, Alexander Iles A, Wilhelm C. Lab on Chip, 11 (2011) 1902. Robert D, Fayol D, Le Visage C, Frasca G, Brulé S, Ménager C, Gazeau F, Letourneur D, Wilhelm C, Biomaterials, 31 (2010) 1586. Chaudeurge A, Wilhelm C, Chen-Tournoux A, Farahmand P, Bellamy V, Autret G, Larghéro J, Thiam R, Desnos M, Hagège A, Gazeau F, Clément O, Menasché P. Cell Transplantation, 21 (2012) 679. Fayol D, Luciani N, Lartigue L, Gazeau F, Wilhelm C. Adv Healthcare Materials, 2 (2013) 312. Di Corato R, Gazeau F, Le Visage C, P Levitz P, Letourneur D, Luciani N, Tillement O, Wilhelm C. ACS nano, In revision. Frasca G, Gazeau F, Wilhelm C. Langmuir. 25 (2009) 2348. Fayol D, Frasca G, Le Visage C, Gazeau F, Luciani N, Wilhelm C. Advanced Materials, In press (2013). Patent B1224FR00. Fayol D, Le Visage C, Ino J, Gazeau F, Letourneur D, Wilhelm C, Cell Transplant Jan 2 (2013).

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[1] Wilhelm C, Gazeau F, Biomaterials, 29 (2008)

381

Recent advances in cell therapy and tissue engineering opened new windows for regenerative medicine, but still necessitate innovative techniques to create and image functional tissues. One promising approach is to associate magnetic nanoparticles with cells in order to supply them with sufficient magnetization to be detectable by MRI [1] or manipulated by magnetic forces [2], while maintaining cell viability and functionalities. A few years ago, we proposed the use of anionic iron oxide nanoparticles as efficient agents for cell internalisation without impacting cell functions [1]. Recently we examined the influence of the amount of internalized iron and the state of nanoparticle aggregation on the capacity for mesenchymal stem cell differentiation and MRI single cell tracking [3]. We then demonstrated that high resolution Magnetic Resonance Imaging (MRI) allowed combining cellular-scale resolution with the ability to detect two cell types simultaneously at any tissue depth [4]. In parallel, we adressed the challenge to create a functional tissue from stem cells in vitro. The aim was to confine stem cells in three dimensions at the millimetric scale by using home-designed miniaturized magnetic devices, in order to create cellular patterns for cartilage tissue engineering [5]. The labeling of endothelial precursors provided as well new possibilities to create potential vessel substitutes [6]. Taken togeter, these results have fundamental implications for the use of magnetic nanoparticles for cell therapy, from MRI imaging studies of stem cell fate to tissue engineering.

Figures

382

Figure 1: From left to right : Magnetic nanoparticles are confined within intracellular endosomes, providing the cell with enough magnetization to be detected individually on high resolution MRI scans - Magnetically driven fusion of stem cell aggregates assembled by micromagnets results in the formation of a continuous tissue layer containing abundant cartilage matrix - A technological approach combining a tubular scaffold and magnetic endothelial cells allows creating a pluricellular and organized vascular graft, with in depth MRI detection of individual cellular components.

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Oral 1

Graphene/graphene oxide – multifunctional platform for drug delivery and photodynamic therapy in cancer treatment

Malgorzata Wojtoniszak

mwojtoniszak@zut.edu.pl

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drug release from graphene oxide in physiological solution – phosphate buffered saline (PBS) containing different biocompatible polymers will be investigated. It was discovered that dispersion of MTX-GO in poly sodium-4-styrene sulfonate (PSS) and poly ethylene glycol (PEG) resulted in significant increase of the release time. The material was characterized with transmission electron microscopy (TEM), atomic force microscopy (AFM), Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR) and UV-vis spectroscopy. Furthermore, antineoplastic action against human breast adenocarcinoma cell line MCF7 of MTX-functionalized graphene oxide and empty graphene oxide was explored. It was found that the anti-proliferative activity of MTX-GO depended on dispersant used to stabilize the suspension.

References [1] H. Park, J.A. Rowehl, K.K. Kim, V. Bulovic, J. Kong, Nanotechnology, 21 (2010) 505204. [2] T. Truong-Huu, K. Chizari, I. Janowska, M.S.

Moldovan, O. Ersen, L.D. Nguyen, M.J. Ledoux, C. Pham-Huu, D. Begin, Catalysis today, 189 (2012) 77-82. [3] Y. Li, G. Zhao, C. Liu, Y. Wang, J. Sun, Y. Gu, Y. Wang, Z. Zeng, Intern. J. Hydrogen Energy, 37 (2012) 5754–5761. [4] H. Wang, T. Chen, S. Wu, X. Chu, R. Yu, Biosensors and Bioelectronics, 34 (2012) 8893. [5] X. Yang, Y. Wang, X. Huang, Y. Ma, Y. Huang, R. Yang, H. Duan, Y. Chen, Journal of materials chemistry, 21 (2011) 3448–3454.

383

Recently, graphene has attracted a great scientific interest in many fields such as electronics, gas storage, catalysis, and medicine among others [14]. Because of its electronic, optical and structural properties it has been explored as a carrier of variety molecules such as drugs for cancer chemotherapy, ferromagnetic for hyperthermia or photosensitizers for photothermal therapy and photodynamic therapy [5-7]. In this study, potential of graphene oxide biomedical applications will be presented. Firstly, in vitro cytocompatibility tests of graphene oxide and reduced graphene oxide dispersed in different polymers will be shown. Here, type of dispersant and concentration of nanomaterials effects on mice fibroblast cells (L929) viability will be reported. Secondly, graphene oxide (GO) functionalization with methylene blue (MB) and its performance in singlet oxygen generation (SOG) under irradiation of laser with excitation wavelengths of 785 nm will be reported. Remarkably, it was noted that GO functionalized with MB (MB-GO) showed an enhanced efficiency in singlet oxygen generation compared to pristine MB. The efficiency in SOG was detected by photobleaching of 9,10anthracenediylbis(methylene)dimalonic acid (ABMDMA). Detailed characterization of the obtained material was carried out with UV-Vis spectroscopy, Raman spectroscopy, FT-IR spectroscopy, and confocal laser scanning microscopy. Interestingly, in contrast to previous study [8], fluorescence confocal microscopy demonstrated that the adsorption of MB on GO resulted in an enhanced fluorescence when the material was excited with light from blue to orange range. Finally, covalent functionalization of graphene oxide with anticancer drug methotrexate (MTX) through amide bonding will be shown. A kinetics of the

NANOBIO&MED 2013

West Pomeranian University of Technology in Szczecin, Institute of Chemical and Environment Engineering, Pulaskiego 10, 70-322, Szczecin, Poland 2 Pomeranian Medical University, Department of General Pathology, Powstańców Wlkp. 72, 70-111, Szczecin, Poland 3 Pomeranian Medical University, Department of Pharmacology, Powstańców Wlkp. 72, 70-111, Szczecin, Poland

D. Rogińska2 M. Perużyńska3 M. Kurzawski3 M. Droździk3 B. Machaliński2 and E. Mijowska1

[6] L.Z. Bai, D.L. Zhao, Y. Xu, J.M. Zhang, Y.L. Gao,

L.Y. Zhao, J.T. Tang, Materials letters, 68 (2012) 399–401. [7] K. Yang, L. Hu, X. Ma, S. Ye, L. Cheng, X. Shi, C. Li, Y. Li, Z. Liu, Advanced materials, 24 (2012) 1868–1872. [8] K. Haubner, J. Murawski, P. Olk, L.M. Eng, C. Ziegler, B. Adolphi, E. Jaehne, Chemphyschem A European Journal Of Chemical Physics And Physical Chemistry, 11 (2010) 2131-2139. Figures

Figure 1: AFM image of graphene oxide functionalized with methotrexate (MTX-GO).

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Figure 2: Fluorescent confocal images of MB-GO obtained in different excitations (blue: A, green: B, orange: C, far red: D).

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Oral Alexey Yashchenok 1 A. Masic 2 D. Gorin B. Parakhonskiy3,4 1 H. Möhwald and 1,5 A. Skirtach

alexey.yashchenok@mpikg.mpg.de

Max Planck Institute of Colloids and Interfaces, Department of Interfaces, Germany Saratov State University, Department of Nano- and Biomedical Technologies, Russia 3 University of Trento, BIOtech center, Italy 4 Shubnikov Institute of Crystallography, Russia 5 Ghent University, Department of Molecular Biotechnology, Belgium 2

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increase of SNR due to SERS allows visualization and detection of biomolecules. The Raman probes developed in this work can be ubiquitously applied for molecular imaging and sensing in cells and tissues. We continue our study for assembling of gold nanoparticles within pores of calcium carbonate microparticles. Porous calcium carbonate microparticles exhibited superior performance in comparison to smooth silica particles. Porosity which can be linked to adsorption of particles is identified as a key mechanism leading to an increased nanoparticle adsorption and enhanced label-free Raman based detection [2]. Using these particles, label-free detection of important biomolecules such as glucose (Fig. 2) and bovine serum albumin is demonstrated at concentrations of nM–μM levels. Such an approach is attractive for developing biomarker sensing at concentrations corresponding to healthy and diseased individuals.

References [1] A. Yashchenok. A. Masic, D. Gorin, B.S. Shim,

N.A. Kotov, P. Fratzl, H. Möhwald, A. Skirtach, Small, 9 (2013) 351. [2] A.M. Yashchenok, D. Borisova, B.V. Parakhonskiy, A. Masic, B.E. Pinchasik, H. Möhwald, and A.G. Skirtach, Annalen der Physik Special Issue: Plasmonic Sensors, 524 (2012) 723.

385

Self-assembly of colloidal noble metal nanoparticles into macroscopic one, two, and three-dimensional arrays either flat or curved substrate is very promising approach, that is to developing sensors based on plasmonic effect. Layer-by-Layer (LbL) deposition is established method for fabrication of multicomponent structures on the solid support in the nanoscale. We applied in this work LbL approach to fabricate nanoplasmonic sensors based on gold nanoparticles adsorbed over carbon nanotubes network and also introduced those into pores of calcium carbonate (CaCO3) microparticles. Roughened structure of developed colloids allowing us to use them as detectors of molecules in Raman experiments based on surface enhancement Raman scattering effect (SERS). Today SERS is one of powerful analytical tool, especially in term of label-free purposes. Compare with fluorescence imaging SERS overcomes photobleaching and provides highly detailed molecular information. Since the discovery of surface enhanced Raman scattering novel platforms have evolved and various approaches to SERS substrates have been introduced. Currently, we achieved remarkable high Raman scattered signal from probe based on silica core coated with carbon nanotubes subsequently functionalized with gold nanoparticles aggregates. Functionalized carbon nanotubes provide easy localization at extremely low laser powers and increased roughness necessary for highly efficient SERS amplification [1]. Upon intracellular incorporation probes significantly enhance molecular fingerprints of biomolecules commonly found inside NIH3T3 fibroblast and enable fast acquisition rates at laser powers completely harmless to living cells (Fig. 1). Remarkable

NANOBIO&MED 2013

Layer-by-Layer assembly of nanocomposite colloidal probes for Raman-based detection of biomolecules

Figures

Figure 1: Raman spectroscopic imaging of a living fi broblast cell with incorporated colloidal probes. (a) Linear combination of the averaged single spectra b) Characteristic for cell compartments (green-cytoplasm, blue-nucleus, yellow-carotenoids) and goldnanoparticle- SWCNT functionalized colloidal probes (red). (c) Intracellular SERS signal recorded at the surface of the probe.

386

Figure 2: SERS spectra of D-Glucose molecules with varying concentration. The characteristic peak at 1130 1/cm is pointed out. All measurements were made using 785 nm laser (10 mW)..

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Oral In vitro diagnostics of microbial cells and their antibiotic resistance using nanostructured anodic aluminum oxide growth platforms 1

Tatiana Zimina 2 L. Kraeva 1 E. Sokolova A. Soloviev1 1 V. Luchinin and 2 G. Hamdulaeva

tmzimina@gmail.com

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functional elements of the system are presented including culture growth platform (Figure 1) based on porous anodic alumina (PAA), AST diffusion and laser speckle optics turbidimetry modules are discussed. The PAA membranes with regular pore arrays and nano porous supports for the cell growth are fabricated using MEMS/MOEMS technologies. These elements could be incorporated into an integrated system for isolation and reliable direct identification of a range of pathogens, as well as express determination of their viability and threshold concentrations of various classes of antibiotics in the “point-of-care” format. The structured colonies growth platform fabricated on the basis of microporous aluminium oxide (AAO) is a principal part of the system1, which could facilitate direct CB method in terms of analysis time, accuracy and point-of-care availability. The main feature of such platform is a possibility to cultivate homogeneous juvenile colonies, sufficient for identification. Using such juvenile colonies accelerates microbiological analysis by factor of 3 and above as well as provides new opportunities for process automation. Isolated small colonies are grown at spatially arranged growth spots as micropores covered with nanoporous AAO. After limited growth time (lower than in conventional microbiological procedures), when colonies achieve a moderate size of approximately 8 to 128 organisms, they should be cautiously displaced. The separation of juvenile colonies from original “nesting sites” is made by microhydroblow generated from underneath by micro pump. In this case micropores are used as jets to form a hydraulic impulse. Then microflow moves them to the next functional elements. This allows to prepare a sample ready for aligning and transport

387

The development of tools for express analysis of human specimens related to aggravated problem of bacteriological threats, combined with the drug resistance is becoming increasingly important. Conventional in vitro microbial cells identification and antibiotic resistance diagnosis is based on culture growth of microorgranisms. Implementation of this direct approach is usually labour intensive, low throughput and slow. The amplification of clinically relevant DNA by the Polymerase Chain Reaction (PCR) and micro arrays provides data on potential pathogenicity while in clinical use it is crucial to have information on resistance of an actual pathogen from a particular clinical case to correctly select medication. Thus, improving culture based (CB) diagnosis methods could contribute to solving the drug resistance problem, via facilitating the optimal and timely choice of medication. Main limiting factors of CB methods, which restrict their express implementation, are: microorganism growth rate (to make a sample available for manual operations), pathogen recognition and determination of its viability in response to antibiotic exposure (antibiotic susceptibility testing - AST). Miniaturization and heterogeneous integration of living bacteria and MEMS components further contribute to resolving the problems associated with classical CB approaches. To produce a miniaturised automated platform, appropriate for integration to carry on total microbiological procedure automatically. The presented devise consists of a number of functional modules, namely: colonies growth platform, microfluidic transport system with liquid key for colonies sorting, express pathogen recognition system, AST growth and viability determination module etc. In this work, essential

NANOBIO&MED 2013

St. Petersburg Electrotechnical University “LETI”, 5 Professora Popova str., St. Petersburg, Russia 2 Pasteur Institute of Epidemiology and Microbiology, 14 Mira str., St. Petersburg, Russia

to other functional elements of total system intended for pathogen identification, sorting and AST. The growth spots are formed in a single technological process of anodic oxidation. The growth platform is intended for further integration targeted at development of full automatic pointof-care microbiological analyser including AST module, with disposable microbial platforms and analysis time 6 – 8 hours. The application of the module in analysis of a number of microbiological samples is described. Acknowledgement: The work was funded by FMBA RF grant No 40.002.12.0 of 20.02.2012 and by the Russian Federation Ministry of Education and Science under Federal Grant Program “Scientific and Educational Human Resources of Innovative Russia”, The State Contracts No 14.B37.21.0568 and No 14B37210793.

References [1] Tatiana M. Zimina1, Victor V. Luchinin1,

Figures

Figure 1: (a) Schematic representation of growth chamber: 1 – AAO substrate, 2 – micropore, 3 – nanoporous Al2O3 membrane permeable for nutrient, 4 – cell, 5 – juvenile colony, 6 – direction of microhydroblow displacing colonies; (b) AAO platform with micropores; (с) the same as (b) with microbial cells; (d) juvenile colony of Staphylococcus aureus, growth time t = 1 h, number of cells N ~ 800, size of colony ~10 microns, (e) juvenile colony of Staphylococcus epidermidis, t = 1 h; (f, g) nanoporous Al2O3, section and top view correspondingly.

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Aleksei V. Soloviev1, Nadezhda V. Kuznetsova1, Nikolai I. Mukhurov2, Irina V. Gasenkova2, Liudmila A. Kraeva3, Galina Ja. Tseneva3, Functional Elements of Laboratoryon-a-Chip for Express Identification and Antimicrobial Susceptibility Testing of Bacterial Respiratory Tract Infections, Proc. of. “Lab-on-Chip”, Dublin, 27-29 May, 2010. [2] T. M. Zimina, A. V. Soloviev, V. V. Luchinin, E. V. Sokolova, A method of microbial cells growth and a device for its realization. Patent app. N 2011154770/10 (082262). Filed 29.12.2011. [3] N. I. Mukhurov, I. V. Gasenkova, I. F. Kotova, E. A. Tyavlovskaya, T. M. Zimina, I. G. Kazarin, N. I. Latnikova, V. V. Luchinin. Nanopore structures of anodic aluminium oxide for analytical Microsystems. Proc. of. “Dielectrics2008”. St. Petersburg. [4] L. Kraeva, G.Tseneva, T.Zimina. Express diagnostics of toxin–positive strains of C. diphtheriae. // Tenth International Meeting of the European Laboratory Working Group on Diphtheria, ELWGD, Larnaca, Cyprus, 5-7 November 2008.

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PPM

2013

Index PPM2013 Contributions Invited

Pag

■ Javier Aizpurua (CFM/CSIC & DIPC, Spain) "The quantum regime in tunneling plasmonics"

399

■ Remi Carminati (ESPCI, France) "Weak and strong coupling to photonic modes in nanoscale disordered media"

407

■ Alfonso Cebollada (IMM-CNM-CSIC, Spain) "Magnetoplasmonics: combining magnetic and plasmonic functionalities"

410

■ Aristide Dogariu (Univ. of Central Florida, USA) “Optical Action in Complex Media”

■ Francisco J. García-Vidal (UAM, Spain) "Theory of strong coupling between quantum emitters and propagating surface plasmons"

424

■ Rainer Hillenbrand (CIC nanoGUNE, Spain) "Infrared Near-field Spectroscopy - From Nanoscale Chemical Identification of Polymers to Real-space Imaging of Graphene Plasmons"

431

■ Tobias Kippenberg (EPFL, Switzerland) "Cavity Optomechanics: Exploring the coupling of light and micro- and nanomechanical oscillators"

433

■ Philippe Lalane (IOGS, Palaiseau, France) "Purcell factor of photonic and plasmonic nanoantennas"

437

■ Maciej Lorenc (IPR- UMR6251, Rennes, France) "Femtosecond Spin-State Photoswitching of Molecular Nanocrystals"

442

■ Lukas Novotny (ETH Zurich, Switzerland) "Cascaded Field Enhancement with Self-Similar Antennas"

444

■ Romain Quidant (ICFO, Spain) "Controlling the interaction of light with the very small"

451

■ Anne Sentenac TBC (Institut Fresnel- UMR 7249, Marseille, France) "Improving the resolution of optical far-field microscopes using structured illumination"

453

■ Niek F. van Hulst (ICFO, Spain) "Non-dipolar & magnetic interactions with optical antennas"

458

■ Martin Wegener (Karlsruhe Inst. of Technology, Germany) "Three-dimensional optical laser lithography: No limits?"

461

■ Diederik Wiersma (LENS, Italy) Abstrat not provided by the speaker

■ Anatoly Zayats (King's College London, UK)

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462

391

"Active Nanoplasmonics: Loss, Gain and Nonlinearity"

Oral

Pag

■ Rodrigo Alcaraz de la Osa (University of Cantabria, Spain) 400

"Quantum effects in small plasmonic particles in the UV-VIS range"

■ Pablo Alonso-González (CIC nanoGUNE, Spain) Visualizing the near-field coupling and interference of bonding and anti-bonding modes in infrared dimer nanoantennas""

402

■ Hidehiko Asanuma (Max Planck Institute of Colloids and Interfaces, Germany) "Molecular self-assembly towards macroscale ordering: application as SERS substrate"

404

■ Alvaro Blanco (ICMM-CSIC, Spain) 405

"Observation and use of internal water in silica photonic structures"

■ Alexandre Cazé (Institut Langevin ESPCI Paristech CNRS, France) 408

"Cross Density of States and spatial coherence in complex plasmonic systems"

■ Alexander Chizhik (Universidad del País Vasco, Spain) "Mechanisms of magnetization reversal in magnetic wires. Magneto-optical study"

411

■ Eduardo Coutino Gonzalez (KULeuven, Belgium) "Luminescent zeolite composites with outstanding external quantum efficiency using silver clusters as dopants"

413

■ Nuno de Sousa (Universidad Autonoma de Madrid, Spain) 415

"Theoretical study of the magneto-optical activity in Au/Co/Au disks"

■ Ruben Esteban (Centro de Fisica de Materiales, Centro Mixto CSIC-UPV/EHU and Donostia International Physics Center (DIPC), Spain)

416

"Linear chains as optical building blocks of complex metallic clusters"

■ Josep Ferré i Borrull (Universitat Rovira i Virgili, Spain) "Simplified Method for 3D-structured Nanoporous Anodic Alumina with stop bands in the visible"

418

■ Luis S. Froufe-Perez (CSIC, Spain) "Wide band transparent metallo-dielectric nanowires at telecommunications wavelengths"

420

■ Juan Galisteo-López (ICMM-CSIC, Spain) 422

"FRET-mediated amplified spontaneous emission in biopolymer complexes"

■ Manuel Gómez (CIQUS, Center for Research in Biological Chemistry and Molecular Materials, Spain) 425

"New SERS substrates made of polymers and Al, cheap and highly efficient"

■ Jordi Gomis Bresco (Institut Català de Nanotechnologia (ICN), Spain) "Optomechanical coupling in 1D corrugated structures with complete dual photonic and phononic band gap"

427

■ Juan Bautista Gonzalez Diaz (CIC nanoGUNE, Spain) "Quantitative Magneto-Optical Characterization of Diffusive Reflected Light from Rough Steel Samples"

428

■ Marcin Grzelczak (Universidad de Vigo, Spain) 430

"Optical response of individual Au-Ag@SiO2 hetero-dimers"

■ Magnus Jonsson (Delft University of Technology, Netherlands) 432

"Nanopores for Bioanalytical Sensing"

■ Oleksandr Kovalenko (Institut des Molécules et Matériaux du Mans, France) 434

"Magnetization switching by ultrashort acoustic pulses"

■ Valentina Krachmalnicoff (Institut Langevin, France)

392

"Nanoscale mapping of the local density of optical states in the near-field of a plasmonic antenna"

436

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Oral

Pag

■ Kristof Lodewijks (Chalmers University of Technology, Sweden) "Designer magnetoplasmonics in ferromagnetic nanoparticles through symmetry breaking"

438

■ Fernando López-Tejeira (Universidad de Zaragoza, Spain) "High Performance Nanosensors Based on Plasmonic Fano-Like Interference"

440

■ Beri N. Mbenkum (Centro Nacional de Microelectrónica (CSIC), Spain) "Size- & Shape-Independent Tuning of Plasmon Resonance Frequency of Sub 10 nm Gold Nanoparticles"

443

■ Vilianne Ntsame Guilengui (Institut d'Electronique du Sud, France) "Localized Surface Plasmons Resonances on GaSb-based materials for Infrared applications"

445

■ Olalla Pérez-González (University of the Basque Country, Spain) "Transport and Sensing in Plexcitonic Nanocavities"

447

■ Mohamed Ameen Poyli (Donostia International Physics Center (DIPC) and Centro de Fisica de Materiales, CFM(CSIC_UPV/EHU)., Spain) "Dielectric Gap-nanoantennas for Low-loss Field Enhanced Spectroscopy"

449

■ Juan Jose Saenz (Universidad Autonoma de Madrid, Spain) "Theoretical and experimental analysis of the directionality of the electromagnetic scattering by magnetodielectric small spherical particles"

452

■ Viktor Shalagatskyi (Institut des Molécules et Matériaux du Mans, France) "The nanostructured acoustic Fresnel lens for focusing THz phonons in gold"

454

■ Francisco Javier Valdivia-Valero (ICMM-CSIC, Spain) "Optical forces on cylinders near subwavelength slits illuminated by either a Gaussian beam or a photonic nanojet: effects of extraordinary transmission and excitation of Mie resonances"

456

■ Alan Vitrey (IMM-CSIC, Spain)

ImagineNano2013

459

393

"Near-field patterns in two dimensional arrays of gold nanorods supporting geometric resonances"

Index PPM2013 Contributions Alphabetical Order I:Invited / O: Oral

Pag

■ Javier Aizpurua (CFM/CSIC & DIPC, Spain) "The quantum regime in tunneling plasmonics"

399

400

402

404

405

407

408

410

411

413

415

416

418

420

422

424

■ Rodrigo Alcaraz de la Osa (University of Cantabria, Spain) "Quantum effects in small plasmonic particles in the UV-VIS range"

■ Pablo Alonso-González (CIC nanoGUNE, Spain) "Visualizing the near-field coupling and interference of bonding and anti-bonding modes in infrared dimer nanoantennas"

■ Hidehiko Asanuma (Max Planck Institute of Colloids and Interfaces, Germany) "Molecular self-assembly towards macroscale ordering: application as SERS substrate"

■ Alvaro Blanco (ICMM-CSIC, Spain) "Observation and use of internal water in silica photonic structures"

■ Remi Carminati (ESPCI, France) "Weak and strong coupling to photonic modes in nanoscale disordered media"

■ Alexandre Cazé (Institut Langevin ESPCI Paristech CNRS, France) "Cross Density of States and spatial coherence in complex plasmonic systems"

■ Alfonso Cebollada (IMM-CNM-CSIC, Spain) "Magnetoplasmonics: combining magnetic and plasmonic functionalities"

■ Alexander Chizhik (Universidad del País Vasco, Spain) "Mechanisms of magnetization reversal in magnetic wires. Magneto-optical study"

■ Eduardo Coutino Gonzalez (KULeuven, Belgium) "Luminescent zeolite composites with outstanding external quantum efficiency using silver clusters as dopants"

■ Nuno de Sousa (Universidad Autonoma de Madrid, Spain) "Theoretical study of the magneto-optical activity in Au/Co/Au disks"

■ Aristide Dogariu (Univ. of Central Florida, USA) “Optical Action in Complex Media”

■ Ruben Esteban (Centro de Fisica de Materiales, Centro Mixto CSIC-UPV/EHU and Donostia International Physics Center (DIPC), Spain) "Linear chains as optical building blocks of complex metallic clusters"

■ Josep Ferré i Borrull (Universitat Rovira i Virgili, Spain) "Simplified Method for 3D-structured Nanoporous Anodic Alumina with stop bands in the visible"

■ Luis S. Froufe-Perez (CSIC, Spain) "Wide band transparent metallo-dielectric nanowires at telecommunications wavelengths"

■ Juan Galisteo-López (ICMM-CSIC, Spain) "FRET-mediated amplified spontaneous emission in biopolymer complexes"

■ Francisco J. García-Vidal (UAM, Spain)

394

"Theory of strong coupling between quantum emitters and propagating surface plasmons"

ImagineNano2013

I:Invited / O: Oral

Pag

■ Manuel Gómez (CIQUS, Center for Research in Biological Chemistry and Molecular Materials, Spain) "New SERS substrates made of polymers and Al, cheap and highly efficient"

425

427

428

430

431

432

433

434

436

437

438

440

442

443

444

445

447

449

451

452

■ Jordi Gomis Bresco (Institut Català de Nanotechnologia (ICN), Spain) "Optomechanical coupling in 1D corrugated structures with complete dual photonic and phononic band gap"

■ Juan Bautista Gonzalez Diaz (CIC nanoGUNE, Spain) "Quantitative Magneto-Optical Characterization of Diffusive Reflected Light from Rough Steel Samples"

■ Marcin Grzelczak (Universidad de Vigo, Spain) "Optical response of individual Au-Ag@SiO2 hetero-dimers"

■ Rainer Hillenbrand (CIC nanoGUNE, Spain) "Infrared Near-field Spectroscopy - From Nanoscale Chemical Identification of Polymers to Real-space Imaging of Graphene Plasmons"

■ Magnus Jonsson (Delft University of Technology, Netherlands) "Nanopores for Bioanalytical Sensing"

■ Tobias Kippenberg (EPFL, Switzerland) "Cavity Optomechanics: Exploring the coupling of light and micro- and nanomechanical oscillators"

■ Oleksandr Kovalenko (Institut des Molécules et Matériaux du Mans, France) "Magnetization switching by ultrashort acoustic pulses"

■ Valentina Krachmalnicoff (Institut Langevin, France) "Nanoscale mapping of the local density of optical states in the near-field of a plasmonic antenna"