Lillie's Method for Nucleic Acids (1965)

Created by Kathleen Patrick, Modified on Mon, 11 Nov at 3:32 PM by Kathleen Patrick

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.