EMS26109

Components:

  • Harris Haematoxylin

  • Weigert's Iron Haematoxylin Solution A

  • Weigert's Iron Haematoxylin Solution B

  • Analine Eosin Solution

  • Naphthol Green B Stain 1%

  • Acid Alcohol 1% In 75% Alcohol

  • Ammonia Water 0.3%

Fixation:

10% Buffered Neutral Formalin

Section:

Paraffin at 6µm

Staining Procedure:  

  1. Deparaffinise and hydrate to distilled water.  Include a control slide with all samples.

  2. Stain in Harris Haematoxylin 10 min. or Weigert's Iron Haematoxylin for 3 min.

  3. To prepare Weigert's Iron Hematoxylin, mix equal parts just before use:
    Weigert's Iron Haematoxylin Sol'n A and Weigert's Iron Haematoxylin Sol'n B.

  4. Wash in running  water for several minutes, differentiate each slide in Acid Alcohol, 1% and then wash again.

  5. Blue each slide in Ammonia Water and wash in running water.

  6. Stain in Aniline-Eosin for 5 minutes, rinse well in distilled water.  Slides should appear deep rose.

  7. Counter-stain in Naphthol Green B for 5 minutes.

  8. Under a microscope, differentiate the slides in 95% alcohol (two changes) until the erythrocytes remain a deep rose.

  9. Dehydrate through two changes absolute alcohol and clear in two changes Xylene.

  10. Mount.

Results:

Erythrocytes (ingested)

rose

Connective tissue

green

Amoebae

blue-green

Nuclei (amoebic)

darker blue-green

References:

Gridley, M.F.:  Am. J. Clin., Pathol., 24:243 (1954).

Luna, L.G.:  Histologic Staining Methods, 3rd ed.:  New York:  McGraw-Hill Book Co., c. 1968, p. 228.