Stool analysis
Lab. Technology
Lab-3-
Microscopic Examination of wet mount
• Wet mount is the simplest and easiest technique for the examination of feces, and
this method should be performed in all laboratories at peripheral level.
• A wet mount can be prepared directly from fecal material or from concentrated
specimens.
• The basic types of wet mount that should be used for each fecal examination are
normal saline (0.85% NaCl), iodine, and buffered methylene blue.
Microscopic Examination of wet mount (cont.)
The Saline Wet Mount
▪ Is used for the initial microscopic examination of stool specimens.
▪ It is employed primarily to demonstrate worms eggs, larvae.
Protozoan trophozoites, and cysts.
▪ This type of mount can also reveal the presence of red blood cells
and white blood cells.
▪ If the presence of amoebic trophozoites is suspected, warm saline
(37˚C) should be used.
Microscopic Examination of wet mount (cont.)
The Iodine Wet Mount
▪ Is used mainly to stain glycogen and the nuclei of cysts, if
present.
▪ Cysts can usually be specifically identified in this mount.
▪ Trophozoite can not be revealed by this type of wet mount,
because iodine kill trophozoite.
Microscopic Examination of wet mount (cont.)
The Buffered Methylene Blue Wet Mount
▪ Should be prepared each time amoebic trophozoites are seen
in a saline wet mount, or when their presence is suspected.
▪ BMB stains amoebic trophozoites, but not stain amoebic cysts, flagellate
trophozoites or flagellate cysts.
▪ BMB stain is appropriate only for fresh unpreserved specimens.
▪ BMB stain live organism only, it isn’t used on preserved samples in which
the organism have been killed
▪ Wait for five minutes to allow the stain to penetrate the trophozoites. It will
overstrain the trophozoites in 30 minutes.
➢Formed stool: take the portion of stool from an area to include inside and
outside parts of the specimen.
➢Stool with mucus: if mucus is present, label a second slide with the
patient’s name or number. Put a drop of saline on the slide, pick up a small
portion of mucus and mix with the saline. Trophozoites, if present, are
sometimes more readily found in mucus than in the solid parts of the stool.
➢Loose watery stool: if mucus is not present, pick up a small portion of the
stool (any part) and mix with the saline.
Making Direct smear Microscopy
➢ Materials and reagents:
✓ Microscopic slides.
✓ Cover slips.
✓ Applicator sticks.
✓ Marker or pen for labeling.
✓ Reagents:
▪ Saline solution (isotonic)
▪ Lugols iodine(1% solution)
▪ BMB
DIRECT SMEAR
A small amount of feces is transferred to a microscope slide and mixed
with saline to produce a layer thin enough to read newsprint through. A
coverslip is placed on the slide for viewing under the microscope.
Rapid screening of fecal material with the ability to observe motile
trophozoites that may be distorted and killed in hypertonic flotation
solutions
Reliability of Results
Dependent on experience and skill of person examining the slide and the
presence and viability of the trophozoites in the small-sample volume.
Wet mount procedure
Examine the slide on microscope:
o 10X
o 40X
Disposal of microscopic slides
• In diluted detergent
• Great care is needed in handling all laboratory specimens and rubber
gloves should always be worn
Result of Examination
• If no parasites are found:
▪ “No ova or parasites seen”, and specify whether this result was
obtained by direct examination or by a concentration method (name
method used).
▪ Never state categorically: “No parasites”
• If any parasites are seen, write the scientific name of the parasite with
stages
• Example: Giardia lambilia cyst
