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 x3 5 min
4. 100% EtOH 5 min
5. 95% EtOH 5 min
6. Protargol-S, 36 C 12-30 hours
7. D-H2O x6 rinse
8. Hydroquinone sol. 15 min
9. D-H 2O x6 rinse
10. Gold Chloride sol. 10-15 min until light gray
11. D-H2O x 6 rinse
12. 2% Oxalic acid 2-3 min until fibers are sharp
13. D-H20 x 6 rinse
14. Hypo soln. 5-10 min
15. D-H20 x 6 rinse
16. van Gieson soln. 1-3 min, opt.
17. 70% EtOH 3-5 min
18. 95% EtOH 3-5 min
19. 100% EtOH x 2 3-5 min ea.
20. Xylene x 2 3-5 min ea.
21. Coverslip