EMS26774
Components:
Schiff's Reagent
Hydrochloric Acid 1N
Sodium Bisulphite Solution 0.05M Aqueous
Fast Green FCF Solution 0.01%
Fixation:
10% Buffered Neutral Formalin
Sections:
Paraffin @ 6µm
Staining Procedures:
- Deparaffinise and hydrate to distilled water.
- Place slides in preheated 1N Hydrochloric Acid at 60°C for 10 -15 minutes.
- Stain for 10 minutes in Schiff’s Reagent.
- Place in Sodium Bisulphite, 0.05M for 3 changes, 2 minutes each. Wash in running water for 5 minutes.
- Stain for few seconds in Fast Green Solution, 0.01%.
- Dehydrate in 96% alcohol, absolute alcohol, and clear in xylene, two changes each.
- Mount with Permount.
Stain Results:
Nuclear Chromatin | Deep Red-Purple |
Cytoplasm | Light Green |
The Chromatin of plasmodia, sarcosporidia toxoplasma and histoplasma | Pale Red |
NOTE: If the fast Green is too intense, it may be removed promptly in water, it does not wash out in alcohol.
References:
Lillie, R.D., Histopathologic Technic and Practical Histochemistry, 3rd ed, McGraw-Hill. NY, pp 149-150, 1965.
Luna, L.G., (ed). Manual of Histologic Staining Methods of the AFIP 3rd edition. McGraw-Hill, NY, p. 131, c. 1968.