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Electron Microscopy Sciences

Technical Data Sheets

Lillie’s Method for Nucleic Acid

EMS Catalog #: 26774-Series

Fixation:

10% Buffered Neutral Formalin (#15741-01).

Sections:

Paraffin @ 6 microns.

Staining Procedures:

  1. Deparaffinize and hydrate to distilled water.
  2. Place slides in preheated 1N Hydrochloric Acid (#26774-02) at 60°C for 10 -15 minutes.
  3. Stain for 10 minutes in Schiff’s Reagent (#26774-01)
  4. Place in Sodium Bisulfite, 0.05M (#26774-03) for 3 changes, 2 minutes each. Wash in running water for 5 minutes.
  5. Stain for few seconds in Fast Green Solution, 0.01% (#26774-04)
  6. Dehydrate in 96% alcohol, absolute alcohol, and clear in xylene, two changes each.
  7. Mount with Permount (#17986-01).

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.

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Lillie’s Method for Nucleic Acid