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Electron Microscopy Sciences

Gridley's Method for Endamoeba Histoslytica

EMS Catalog #: 26109-series

Fixation:

10% Buffered Neutral Formalin (#15740)

Section:

Paraffin at 6 microns

Staining Procedure :  

  1. Deparaffinize and hydrate to distilled water.  Include a control slide with all samples.
  2. Stain in Harris Hematoxylin, (#26109-01), 10 min. or, Weigert's Iron Hematoxylin, 3 min.
    1. To prepare Weigert's Iron Hematoxylin, mix equal parts just before use:
      Weigert's Iron Hematoxylin Sol'n A, (#26109-1A) and
      Weigert's Iron Hematoxylin Sol'n B, (#26109-1B).
  3. Wash in running  water for several minutes, differentiate each slide in Acid Alcohol, 1%, (#26109-04), and then wash again.
  4. Blue each slide in Ammonia Water, (#26109-05), wash in running water.
  5. Stain in Aniline-Eosin, (#26109-02), 5 minutes, rinse well in distilled water.  Slides should appear deep rose.
  6. Counter-stain in Naphthol Green B, (#26109-03), 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 (#23400).
  9. 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.

Online Ordering

Gridley's Method for Endamoeba Histoslytica is available online from the EMS Catalog. For ordering or product information, click here.

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