10X Phosphate Buffered Saline (PBS): To prepare 1 liter, Na2HPO4 --------------------- 10.9 g NaH2PO4 --------------------- 3.2 g NaCl --------------------------- 90 g Distilled water -------------- 1000 ml Mix to dissolve and adjust pH to 7.4 0.2M Citric Acid: To prepare 100 ml, Citric acid (monohydrate) ------ 4.2 g Distilled water -------------------- 100 ml 0.2M Sodium Citrate Buffer (fresh): To prepare 100 ml, Sodium citrate (dihydrate) ------ 5.88 g Distilled water --------------------- 100 ml Adjust pH to 7.4 with 0.2M citric acid (keeps refrigerated). Fixative (4% Paraformaldehyde and 0.5% Glutaraldehyde in 0.1M PB): To prepare 1 liter, Paraformaldedyde -------------- 40 g 0.1M PB --------------------------- 1000 ml Heat to 65 C. Add a few drops of 1N NaOH to clear the solution. Stir to dissolve. Filter the solution and add 10 ml of 50% glutaraldehyde. Post-fixative (1% Osmium Tetroxide in 0.1M PB): To prepare 20 ml, 4% osmium tetroxide ---------- 5 ml 0.2M PB --------------------------- 5 ml 0.1M PB -------------------------- 10 ml 2% Glutaraldehyde in PBS: To prepare 100 ml, 50% glutaraldyhyde ------------ 4 ml PBS -------------------------------- 100 ml |