Grocott’s Methanamine Silver (GMS)

Here is another fungus stain.  Some of the fungi that you might stain with PAS but do not will stain with the GMS.  The silver used must be disposed of properly (not down the sink).  When using gold chloride remember to use plastic forceps, metal forceps will corrode on contact.  All glassware should be chemically cleaned before use to reduce contamination.  Pictures will be coming soon.

Use Control Tissue with known fungi.

Safety equipment: Work under a hood with lab coat, gloves, and glasses.

  1. De-paraffin slides in xylene (1) —————— 2 minutes (re-use)
  2. De-paraffin slides in xylene (2)—————— 2 minutes (re-use)
  3. Clear slides in 100% alcohol——————— 2 minutes
  4. Clear slides in 100% alcohol——————— 2 minutes
  5. Hydrate slides in 95% alcohol——————- 2 minutes
  6. Hydrate slides in running water—————– 5 minutes
  7. Place in 10% chromic acid ———————- 15 minutes (re-use)
  8. Rinse in running water ————————- 1 minute
  9. Place in 1% sodium bisulfate ——————– 1 minute (re-use)
  10. Rinse in running water ————————- 1 minute
  11. Place in working Methanamine silver at 60C—- 60 minutes
  12. Rinse in running water ————————- 1 minute
  13. Place in 0.1% gold chloride ——————— 4 minutes (re-use)
  14. Rinse in running water ———————— 1 minute
  15. Place in 2% sodium thiosulfate —————- 4 minutes (re-use)
  16. Rinse in running water ———————–  1 minute
  17. Place in working light green ——————  30 seconds
  18. Dehydrate in 95% alcohol——————— 30 seconds
  19. Dehydrate in 100% alcohol——————– 1 minute
  20. Dehydrate in 100% alcohol——————– 2 minutes
  21. Dehydrate in 100% alcohol——————– 2 minutes
  22. Clear in xylene (3)—————————- 2 minutes (re-use)
  1. 23.   Clear in xylene (4)——————————– 5 minutes (re-use)

Results:         Fungi – Sharply delineated in Black      

Mucin – Taupe to dark Grey

                       Inner parts of mycelia and hyphae – Old Rose            

Background – Pale Green

 

An Expert Histology Service


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