特殊染色
BODIAN STAIN
SOLUTIONS:
1.Protargol-S: 1%; sprinkle Protargol-S on surface of DH2O in beaker, allow granules to dissolve in dark without stirring or heat; pour into staining dish containing 5% copper shot to prevent tissue from being impregnated with silver.
2.Hydroquinone solution: must be made fresh; mix I g hydroquin-one with 5 ml 40% formaldehyde in 100 ml D-H2O.
3.0.2% Gold Chloride
4.2% Oxalic acid
5.5% Sodium thiosulfate (Hypo)
6.Van Geison';s solution: 2.5 ml of 1% acid fuchsin in 97.5 ml saturated aqueous picric acid.
PROCEDURE:
1. Fix tissue with 10% neutral buffered formalin (pH 6.8-7.0) or Bouin';s solution.
2. Section.paraffin blocks 4-5 urn.
3. Deparaffinize in Xylene x35 min
4. 100% EtOH5 min
5. 95% EtOH5 min
6. Protargol-S, 36 C 12-30 hours
7. D-H2O x6rinse
8. Hydroquinone sol.15 min
9. D-H 2O x6rinse
10. Gold Chloride sol.10-15 min until light gray
11. D-H2O x 6 rinse
12. 2% Oxalic acid2-3 min until fibers are sharp
13. D-H20 x 6rinse
14. Hypo soln. 5-10 min
15. D-H20 x 6rinse
16. van Gieson soln.1-3 min, opt.
17. 70% EtOH3-5 min
18. 95% EtOH3-5 min
19. 100% EtOH x 23-5 min ea.
20. Xylene x 23-5 min ea.
21. Coverslip