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EMS Technical Data Sheets
Gridley's Method for Endamoeba Histoslytica
EMS Catalog #: 26109-series
Fixation:
10% Buffered Neutral Formalin (#15740)
Section:
Paraffin at 6 microns
Staining Procedure :
- Deparaffinize and hydrate to distilled water. Include a control slide with all samples.
- Stain in Harris Hematoxylin, (#26109-01), 10 min. or, Weigert's Iron Hematoxylin, 3 min.
- 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).
- To prepare Weigert's Iron Hematoxylin, mix equal parts just before use:
- Wash in running water for several minutes, differentiate each slide in Acid Alcohol, 1%, (#26109-04), and then wash again.
- Blue each slide in Ammonia Water, (#26109-05), wash in running water.
- Stain in Aniline-Eosin, (#26109-02), 5 minutes, rinse well in distilled water. Slides should appear deep rose.
- Counter-stain in Naphthol Green B, (#26109-03), 5 minutes.
- Under a microscope, differentiate the slides in 95% alcohol (two changes) until the erythrocytes remain a deep rose.
- Dehydrate through two changes absolute alcohol and clear in two changes Xylene (#23400).
- 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.