Modified Trichrome Stain Kit (For Microsporidia Spores In Stool And Duodenal Aspirates)
Solution II - Methanol Fixative
Solution III - Modified Trichrome Stain
Solution IV - Acid Alcohol
Solution V - 95% Alcohol
Solution VI - 100% Alcohol
Solution VII - Xylene
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.
ENG Scientific, Inc., 82 Industrial East, Clifton, NJ 07012. Tel: 973-472-7200; Fax: 973-472-9460.