EMS26252

Components:

  • Mayer's Haematoxylin

  • Eosin Y 0.1% Aqueous

  • Azure II 0.1% Aqueous

  • Phosphate Buffer 0.1M, pH 6.8-7.0

Fixation:

Zenker’s / Formalin

Sections:

Paraffin at 6µm

Staining Procedure:              

  1. Deparaffinise and hydrate to distilled water.

  2. Mayer’s Haematoxylin for 15 minutes. Wash in tepid tap water for 15 minutes.

  3. Stain in working solution overnight. Prepare working solution fresh each time as follows:

    1. Eosin Y, 0.1%, Aqueous - 10ml
      Triple distilled water or Phosphate Buffer - 100ml
      Azur II, 0.1%, Aqueous - 10ml
       Glacial Acetic Acid - 2 drops

  4. Differentiate in 95% alcohol until blue ceases to come out into alcohol and erythrocytes and collagen are pink.

  5. Dehydrate in absolute alcohol, clear in xylene, two changes each.

  6. Mount with Permount.

Stain Results:

Nuclei

Blue

Basophile leucocyte, mast cell

Purple to Violet

Cartilage

Purple

Erythrocytes, eosinophile granules

Pink

Cytoplasm

Blue to Pink

References:

AFIP, Manual of Histologic Staining Methods, 3rd edition, Ed. L. Luna: New York: McGraw Hill Publications, c. 1968, p. 120

Mallory, F.B., Pathological Technique, New York, Hafner Publishing Co., 1961, p. 196.