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The blocking and antibody solutions are pipetted directly on the tissue section and covered with a small coverslip made by cutting a section of Parafilm. The coverslips are removed from the slide by immersing the slide in PBST in a vertical staining jar. Slides are washed in a Wheaton staining dish on a rocking platform agitator such as a VariMix platform (Thermolyne). 28. The new solution pushes the previous solution down out of the capillary gap.
Acknowledgments We are grateful to Christian Young for excellent technical assistance and suggestions. The work of the authors was supported by Canadian Institutes of Health Research grants 82790 and 102606, a Canada Research Chair on Hypertension and Vascular Research from Canadian Institutes of Health Research/Government of Canada, and the Canada Fund for Innovation (all to E.L.S.). K.K.M is supported by CIHR grant 115000 and is a Junior 2 Chercheur Boursier of the FRQ-S. References 1. Schiffrin EL (2008) The flame that lights the fire: oxidative stress, inflammation, and renal damage in angiotensin II-induced hypertension. Hypertension 52:205–206 2. Schiffrin EL (2010) T lymphocytes: a role in hypertension? Curr Opin Nephrol Hypertens 19:181–186 3. Schiffrin EL (2013) The immune system: role in hypertension. Can J Cardiol 29(5):543–548 4. Barhoumi T, Kasal DA, Li MW, Shbat L, Laurant P, Neves MF, Paradis P, Schiffrin EL (2011) T Regulatory lymphocytes prevent angiotensin II-induced hypertension and vascular injury. Hypertension 57:469– 476 5. De Ciuceis C, Amiri F, Brassard P, Endemann DH, Touyz RM, Schiffrin EL (2005) Reduced vascular remodeling, endothelial dysfunction, and oxidative stress in resistance arteries of angiotensin II-infused macrophage colony- stimulating factor-deficient mice: evidence for a role in inflammation in angiotensininduced vascular injury. Arterioscler Thromb Vasc Biol 25:2106–2113 6. Guzik TJ, Hoch NE, Brown KA, McCann LA, Rahman A, Dikalov S, Goronzy J, Weyand C, Harrison DG (2007) Role of the T cell in the genesis of angiotensin II induced hypertension and vascular dysfunction. J Exp Med 204:2449–2460
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