Staining procedure

Set up 10 staining (Coplin) jars as shown in Figure 33.1.

1 70% ethanol: 30min to harden and fix smear.

2 70% ethanol: 10min.

4 70% ethanol: 3-5min. Slides may remain here up to 24h.

Figure 33.1 Set up of staining jars for trichrome stain for intestinal protozoa. Change contents of all jars except those with trichrome stain and Histoclear when about 10 slides have been processed. Change trichrome stain and Histoclear about every 50 slides or more often depending on the staining of the positive control.

Figure 33.1 Set up of staining jars for trichrome stain for intestinal protozoa. Change contents of all jars except those with trichrome stain and Histoclear when about 10 slides have been processed. Change trichrome stain and Histoclear about every 50 slides or more often depending on the staining of the positive control.

5 Trichrome stain: 6-8min, remove excess stain with absorbent tissue.

6 Acid-alcohol: 5-10s; transfer to jar 7 immediately stain runs from smear.

7 95% ethanol: two dips. Change ethanol frequently.

8 95% ethanol: 5min.

9 Absolute ethanol: 3min.

10 Histoclear: 3min.

11 Mount the slide with DPX under a coverglass. Allow to dry.

Examine preparation under the light microscope using 10*, 40*, and oil immersion objectives. Protozoal cysts and trophozoites are stained in different shades of red and blue-green against a green background. Chromidial bars are red.

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