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


Staining

  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.


Staining 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.