This stain method was designed as a simple, non-invasive technique for the rapid detection of microsporidia spores in fecal material by light microscopy. The chromotrope based stain is a modification of the trichrome stain for intestinal protozoa.
RECOMMENDED PROCEDURE: Specimens for light microscopial examination should be prepared from a suspension of unconcentrated liquid stool (10 ul) in Sol. I (1 :3 ratio). Smears are prepared directly and thinly spread on a glass slide covering a 45 x 25mm area. Fix smears in Sol. II for 5 minutes. Stain smears with Sol. III for 90 minutes. Rinse slides in Sol. 1V for 10 seconds. Rinse briefly in Sol. V to remove acid. Dehydrate in Sol. V for 5 minutes, Sol. VI for 10 minutes and Sol. VII for 10 minutes. Read microscopically under oil immersion lens (magnification x 1000).
RESULTS: Enterocytozoon bienuesi spores appear ovoid and refractile at approximately 1.5 by 0.90 um. The spore wall of properly stained smears should appear a bright pinkish red. Most bacteria and background debris stain a faint green.
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