EMS26758

Components:

  • Weigert's Iron Haematoxylin A

  • Weigert's Iron Haematoxylin B

  • Fast Green FCF Solution, 0.02%

  • Bismark Brown Y Solution, 0.1%

  • Safranin O, 0.1%

  • Acetic Acid, 1% Aqueous

Fixation:

Formalin, Bouin's, Carnoy, Orth's or Zenker's

Sections:

Paraffin

Procedure:

  1. Deparaffinise and hydrate to distilled water.

  2. Stain in working Weigert's Solution* for 6 minutes. Wash in water. *Prepare working solution by mixing equal volumes of Weigert's Iron Haematoxylin A and Weigert's Iron Haematoxylin B.

  3. Stain in 0.02% Fast Green FCF for 3 minutes.

  4. Wash in 1% Acetic Acid.

  5. Stain for 4-6 minutes in 0.1% Bismarck Brown or in 0.1% Safranin O.

  6. Dehydrate with 95% alcohol and 2 changes of absolute alcohol.

  7. Clear with Alcohol: Xylene, 1:1, 1 change and two changes of Xylene.

  8. Mount with a synthetic resin.

Stain Results:

Nuclei

Black

Cytoplasm

Grey-green

Mucus, cartilage and mast cell granules

Yellowish brown, deep red or orange -red depending on dye used.

References:

Weigert, K., Z. Wissen. Mikr., 21:1-5, 1904.

Lillie, R.D., Histopathologic Technic and Practical Histochemistry , 3 rd , edition, McGraw-Hill , NY , 1965.

Clark , G., (ed.), Staining Procedures , 3 rd edition, Williams & Wilkins, Baltimore , p. 40, c 1973.