EMS26312


Components:

Coleman's Feulgen

Light Green Stock Solution 0.2%

Periodic Acid 0.5% Aqueous

Acid-Alcohol 1%

Ammonia Water 0.3%

Schiff'S Reagent

Light Green Working Solution

Harris Haematoxylin


Fixation:

10% Buffered Neutral Formalin or Zenker’s


Sections:

Cut Paraffin, 6µm


Staining Procedures:

  1. Deparaffinise and hydrate to distilled water.
  2. Oxidise in Periodic Acid, 0.5%, 5 minutes and rinse in distilled water.
  3. Stain in Coleman’s Feulgen or Schiff’s Reagent, 15 minutes and wash in running water to develop the pink colour, 10 minutes.
  4. Counterstain in Harris Hematoxylin 6 minutes, or Light Green Working Solution 5 – 10 seconds. Light Green is better used when delineation of fungi is required. Prepare the working solution by diluting Light Green, 0.2% Solution 1:5 ratio with distilled water; proceed to step #7, dehydration. Tap water and ammonia decolourise Light Green.
  5. Wash in running water and transfer to Acid Alcohol, 1%, for 3 -10 quick dips. Wash again in distilled water.
  6. Dip in dilute Ammonia Water, 0.3%, to blue the sections and again wash in running water for ten minutes.
  7. Dehydrate in 95% alcohol, absolute alcohol, and clear in xylene, two changes each.
  8. Mount with Permount or DPX.


Stain Results:

Nuclei

Blue

Fungi

Red

Background, when Light Green is used as the counterstain

Green, pale


References:

McManus, JFA: Stain Tech.; 23:99 (1948).

AFIP, Manual of Histologic Staining Techniques; L.G. Luna, 3rd edition, New York: McGraw-Hill Publication, c. 1968, p. 160.

Mowry, R.W.: Annals of the New York Academy of Science; 106: 402 (1963).