Gridley's Method for Endamoeba Histoslytica

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.
   1. 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.
3. Wash in running  water for several minutes, differentiate each slide in Acid Alcohol, 1% and then wash again.
4. Blue each slide in Ammonia Water and wash in running water.
5. Stain in Aniline-Eosin for 5 minutes, rinse well in distilled water.  Slides should appear deep rose.
6. Counter-stain in Naphthol Green B for 5 minutes.
7. Under a microscope, differentiate the slides in 95% alcohol (two changes) until the erythrocytes remain a deep rose.
8. Dehydrate through two changes absolute alcohol and clear in two changes Xylene.
9. Mount.

Results:

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.