EMS26364


Components:

Phosphomolybdic/Phosphotungstic Acid Solution

Weigert's Iron Haematoxylin A

Weigert's Iron Haematoxylin B

Biebrich Scarlet Solution 1%

Fast Green FCF Solution 2.5%

Acetic Acid 1% Aqueous

Lugol's Iodine

Sodium Thiosulfate 5% Aqueous

Gram's Iodine Solution


Fixation:

Bouin’s or Russell’s Zenker -contain no Mercury


Sections:

Paraffin at 6µm


Staining Procedures:

  1. Deparaffinise and hydrate to distilled water.
  2. If Zenker’s contain Mercury is used as fixative, remove Mercuric Chloride Crystal by placing in Lugol’s Iodine or Gram’s Iodine for 15 minutes. Rinse in water and clear in Sodium Thiosulfate, 5% for 3 minutes. Wash in water for 10 minutes or longer. If Russell’s Zenker is used, go to step #3.
  3. Stain in Weigert’s Iron Hematoxylin Working* solution for 5 minutes. Wash in running water.
    1. * To prepare working solution, mix equal parts of Weigert’s Iron Haematoxylin A and Weigert’s Iron Haematoxylin B.
  4. Stain in Biebrich Scarlet Solution for 4 minutes. Rinse in water
  5. Mordant in Phosphomolybdic Acid-Phosphotungstic Acid for 1 minute.
  6. Stain in Fast Green FCF Solution for 4 minutes.
  7. Differentiate in Acetic Acid, 1% for 1 minute.
  8. Carry through alcohol and dehydrate with acetone or absolute alcohol.
  9. Clear and mount in balsam saturated with salicylic acid.


Stain Results:

Nuclei

Black

Cytoplasm

Brown to Pink

Muscle

Red

Erythrocytes

Brilliant Scarlet

Collagen

Blue to Blue-Green

Myelinated Fibres

Red

Fibrin

Red to Pink


References:

Clark, G.: Staining Procedures. Williams and Wilkins Co., Baltimore, 3rd ed., c. 1973, p. 59

Lillie, R.D., Arch. Path., 29:207, 1940.