STOOL

EXAMINATION
A COMPREHENSIVE APPROACH

Presented by: Lubna

Macroscopic Examination
Colour

of the specimen.
Texture: formed, semi-formed,
unformed or fluid
Presence of blood, pus, mucus,
worms, or tape worm segments?
Normal adult faeces appear brown
and formed or semi-formed.
Infant faeces are yellow, green and
semi-formed.

Appearance

Possible cause

Unformed containing pus and mucus mixed with Shigellosis , Campylobacter enteritis
blood.
Unformed with blood and mucus (acid pH)

Amoebic dysentery

Unformed or semi formed often with blood and Schistosomiasis

mucus
Watery stool

ETEC infection, Rotavirus enteritis

Rice water with mucous flakes

Cholera

Unformed or watery and sometimes with blood Salmonella infection

and pus.
Unformed, pale colored, frothy, unpleasant Giardiasis, Other conditions that cause malabsorption
smelling stools that floats on water (High fat e.g. post infective tropical malabsorption.
contents)

Fluid stools (containing lactose) with pH below Lactase deficiency

6.
Unformed or semi-formed black stools
(Positive occult blood)

Melaena, Hookworm disease, Iron therapy

Microscopic Examination:
A. Direct Microscopy:
Label a frosted end slide with Patient ID
Place a drop of saline at one end and a drop of
iodine at the other end of the slide.
Using a wooden applicator stick, emulsify a
portion of specimen firstly in the saline and
then in the iodine.
Apply a cover slip to each preparations.
The preparations should be thick enough so as
to just read newsprint through them.
Examine both wet mounts for the presence of
leucocytes, erythrocytes, cysts, ova
Report the results into a log.

B. Stool Concentration Technique:

a) Sedimentation: Formol ether sedimentation.
b) Floatation:
Simple Floatation using Saline
ZnSO4 centrifugal floatation
Procedure for Formol Ether Sedimentation Technique
) Add 9ml of 5% - 10% formalin to the flat-bottomed tube.
) Add 3 rounded spoonful of preserved (or 1 spoonful of fresh)
stool.
) For fresh stool specimens, allow 30 minutes for fixation
before proceeding to the next step.
) Add 3 drops of Triton X-100 to the mixed specimens.
) Add 3ml of ethyl acetate.
) Pull the vent-straw in the strainer unit out approximately 1
inch.

 Attach the FPC strainer tightly to the flatbottomed tube containing the faecal specimen.
Shake vigorously for 30 seconds.
Pointing the conical end downward, shake the
specimen through the strainer into the 15 ml
centrifuge tube.
Unscrew the FPC strainer with the flat-bottomed
tube still attached.
Discard the transport tube and strainer in an
appropriate manner.
Cap the 15ml tube and centrifuge at 500 x g for
10 minutes.

After centrifugation, the specimen should appear clearly

separated into four layers. See illustration below:

 Rim the debris layer using an applicator

stick. Pour off the debris and supernatant
fluid.
With the tube still inverted, use a cottontipped applicator stick to clean and remove
the remaining debris and ethyl acetate.
Return the tube to an upright position and
add 2 to 3 drops of 5% to 10% formalin,
saline and mix the sediment thoroughly.
Prepare slides with a disposable transfer
pipette, Coverslip and examine.
Use the remaining specimen to examine for
coccidian oocysts.

 Scan the entire Coverslip area systematically using

10 x objectives.
Use the 40x objective to examine at least one third of
the coverslip area, as well as any areas where
suspicious objects may have been seen on LP.
Following low power scanning, adding iodine may
facilitate the detection of cysts by enhancing
morphological detail. Use the 40x objective for this
examination.
Adding iodine prior to low power scanning should be
done cautiously; at lower magnification, some
helminth eggs may stain darkly as to appear to be
faecal debris.
Formol ether sedimentation can be used for
detection of all helminthic and protozoan ova and
cysts.

RELATIVE SIZE OF PARASITES

C. Permanent Staining
Proto-fix Wheatleys Trichrome Stain Set is
used for this purpose.
Rapid staining procedure for intestinal
amoeba and flagellates.

Specimen Collection and

Preparation

Permanent smears can be prepared from

Unconcentrated specimen fixed in Proto-fix
A specimen in Proto-fix hat has been concentrated
using CONSED concentration procedure.

Preparation of smear:
Transfer 1-2 drops of the Proto-fix fixed specimen to a
slide.
May use coated slides like CELLBOND for better
adhesion.
Lay or hold the slide flat with specimen side up.
Gently and evenly spread the sample.
Spread the sample out to create thick and thin areas
using a chopping motion.
Allow to stand for 1-2 minutes. Or until it is dried.

Staining Procedure
70% Ethanol - 1.5 minutes. Drain off excess.
70% Ethanol - 1.5 minutes. Drain off excess.
Trichrome stain13 minutes. Drain off excess.
90% Acid Ethanol- 1 3 seconds. Drain off excess.
100% Ethanol 5 10 seconds. Drain off excess.
100% Ethanol - 1 minutes. Drain off excess.
100% Ethanol - 1 minutes. Drain off excess.
PRO-Clear - 3 minutes. Drain off excess.
Mount Cover Slip and examine under Oil Immersion
Objective.

Expected Results
Nuclear Chromatin, Chromatoid Bodies,
Ingested RBCs, Bacteria- Purple to red-violet
Cytoplasm of trophozoites and cysts- Blue
tinted with Purple
Macrophages, WBCs, Yeast- Variable- Green
to blue to purple or red
Background Material and artifacts bluegreen to purple

MICROMETRY

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