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Satoru Eguchi and Katherine Elliott
2.6 Sirius Red Staining
1. Paraffin-embedded tissue sections mounted on slides. 2. Xylene. 3. Ethanol. 4. Weigert’s iron hematoxylin A solution. 5. Weigart’s iron hematoxylin B solution. 6. 0.1 % Sirius Red in saturated picric acid. 7. 0.01 N HCl.
2.7 Masson’s Trichrome Staining
1. Paraffin-embedded tissue sections mounted on slides. 2. Xylene. 3. Ethanol. 4. Weigert’s iron hematoxylin A solution. 5. Weigart’s iron hematoxylin B solution. 6. Bouin’s solution. 7. Biebrich scarlet-acid fuchsin solution. 8. Phosphotungstic-phosphomolybdic acid solution. 9. Aniline blue solution. 10. 1 % acetic acid.
3 Methods 3.1 Osmotic Minipump Implantation
Fill micro-osmotic pump 1. Weigh the empty pump and flow moderator, together (see Note 1). 2. Prepare stock solution of AngII (see Note 2). Taking into account the animal’s weight, calculate dilution of stock solution so that 1000 ng/kg/min is delivered to the animal for 2 weeks.
(a) 140{[10/(1000 × body weight/1 × 106/0.25 × 60)] − 1}.
3. Draw the solution of Ang II into a 1 ml syringe. Attach the 27 G filling tube (see Note 3). 4. Remove the flow moderator from the pump and hold the pump upright. Insert the filling tube as far as possible into the opening at the top of the pump. 5. Inject drug slowly into pump. Remove tube when solution reaches the outlet. 6. Wipe excess solution and insert flow moderator until the white flange is flush with the top of the pump. 7. Weigh the filled pump to determine the weight of the solution loaded (see Note 4).
