The staining technique described here has an advantage that there is minimal cell distortion and the early stages are clearly differentiated. Thus, this stain is superior to earlier described, because it shows stages in encystment as well as the mature cyst.
The method is as follows. Bacteria from solid or liquid media are suspended in the staining solution for wet mount preparation. The staining reagent can be prepared by adding the following ingredients in 100 ml of water. Glacial acetic acid, 8.5 ml; sodium sulphate (anhydrous), 3.25 g; neutral red, 200 mg; light green S.F. yellowish, 200 mg and ethanol, 50ml. After 15 minutes of incubation, remove the amorphous precipitate by filtering through 0.5 μ membrane filter
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