Histological Staining Techniques

Gomori's hexamine silver (methenamine silver) for basement membranes
Gomori, 1946; Jones, 1957.
No gelatine or poly-l-lysine coated slides must be used. Use normal kidney as a control.
  • Dewax sections, rinse in alcohol, rinse in water.
  • Rinse in distilled water.
  • Treat in 1% periodic acid - 10 minutes. Periodic acid solution should be less than 1 month old.
  • Wash well in several changes of distilled water.
  • Place in preheated hexamine silver solution at 56oC.  Rinse all sections in distilled water and examine after 20 minutes. Basement membranes should be blackened, look at the glomeruli in the control section. This step may have to be extended to a maximum of 45 minutes.
  • Wash well in several changes of distilled water.
  • Tone in 0.1% gold chloride (sodium tetrachloroaurate) - 2 minutes.
  • Rinse in distilled water.
  • Fix in 5% sodium thiosulphate - 5 minutes.
  • Wash in water.
  • Counterstain in 0.2% light green in 0.2% acetic acid - 1 minute.
  • Rinse in tap water.
  • Dehydrate, clear and mount.

Results: 


Basement membranes - black.
Background - green.

Stock solution (hexamine silver)
5mls of 5% aqueous silver nitrate- 100mls of 3% aqueous hexamine. Store at 4oC, keeps for 1-2 months.

Working solution
Dilute 2mls of 5% borax with 25mls of distilled water, mix and add 25mls of stock solution.

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