PARASITOLOGY LABORATORY DISCUSSION DISPOSAL OF MICROSCOPE SLIDES

JANUARY 27,2023 Slides should be discarded into pot containing 1%

hypochlorite solution or buried in disposable specimen
containers if they are not to be cleaned for reuse.
LABORATORY SAFETY

As laboratory personnel, laboratory safety practice must

PARTS OF MICROSCOPE
be observed, and it includes the following:
MICROSCOPES are instruments that are used in science
 Proper PPE
laboratories, to visualize very minute objects such as cells,
 Handwashing after work of infectious materials
microorganism, giving a contrasting image, that is
 Disinfection of the material immediately after use magnified.
 Disinfection of contaminated waste
 Properly disposing contaminated waste 1. EYEPIECE/ OCULAR LENS- the lens we look
 Universal Precautions to protect health-care through, usually 10-15x magnification.
workers from exposure to bloodborne pathogen, 2. EYEPIECE TUBE- it holds the eyepiece and
primarily Hepatitis B and HIV connect the eyepiece with the objective lens.
-all patients are assumed to be possible carriers 3. OBJECTIVE LENSES- these are the main lenses,
of bloodborne pathogens help for the visualization of specimen. These
lenses are closest to the object (specimen). A
microscope usually contains 3 or 4 objective
HANDLING OF SPECIMEN lenses, these are 4X (shortest lens), 10X, 40X, and
100X (longest lens). All high-quality microscopes
Great care is needed in handling all laboratory specimen contain achromatic, par centered, parfocal lenses.
and rubber gloves should be always worn. 4. ADJUSTMENT KNOBS- These knobs are used to
BLOOD SAMPLES- must be regarded a potentially focus the microscope on object. There are mainly
infectious, a great care is required when collecting and two different types of adjustment knobs such as
processing sample. fine adjustment knobs and coarse adjustment
knobs.
-Particular risk are: 5. NOSE PIECE/ REVOLVING TURRET- It is a circular
structure in which all the objective lenses are
 Stabbing and Cutting Injuries- dispose of used
screwed. It is used to change the magnification
needles or lancets in a puncture proof container
power, by simply rotate the turret.
or buried in a disposable specimen container
6. THE STAGE- it is a flat platform, which support
after soaking in a disinfectant. Do not reuse
the slides. The specimen placed over this section
lancets and needles. Do not leave used lancets
for viewing. It contains STAGE CLIPS which hold
lying around the laboratory.
the specimen slides firmly in place. Some modern
 Contamination of damaged skin or mucous
microscopes contain MECHANICAL STAGE with
membranes- cover any cuts with impervious
adjustment knobs that allows for more precise
dressing. Avoid spilling of blood into the skin or
positioning of the slides.
mucous membranes. Pipetting by mouth should
7. THE APENTURE- a hole on the microscope stage.
be absolutely forbidden. If blood is spill into the
The light transmitted through this hole from the
skin, immediately wash with soap and water.
source of specimen.
STOOL SAMPLES- skin contact must be avoided. Sample 8. ILLUMINATOR- basically a light source for a
should be soaked in disinfectant solution and then out in microscope found at the base.
a disposable specimen container. 9. CONDENSER LENS- this lens help focus the light
from the source into the specimen and allows to
URINE SAMPLE- skin contact must be avoided. Samples produce sharp images at magnifications of 400x
can be discarded via the sewage system. and above.
10. DIAPHRAGM/ IRIS- located under the stage of a  Among the specimen available for parasitic
microscope and its main function is to regulate examinations, the stool is mostly commonly
the amount of light that strikes the specimen. utilized.

EXAMINATION OF STOOL OR FECAL SAMPLE

 The most common method of diagnosis of

intestinal parasite is through the demonstration
of eggs, larvae, adults, trophozoites, cysts, or
oocysts in the stool.
 The fecal specimen is best collected in clean,
wide-mouthed containers made of waxed
cardboard or plastic with a tight-fitting lid to
ensure retention of moisture and to prevent
accidental spillage.

METHODS OF EXAMINATION

 Stools samples are submitted to the

laboratory in the fresh state or preserved
samples.
 If the stool is fresh, the laboratory can classify
consistency of the stools as formed, semi-
formed, soft, loose, or watery.
 The color of the stool can be indicative of the
LABORATORY DIAGNOSIS presence of the parasite.
 By gross examination of the stools, tapeworm
Most parasitic disease cannot be established based
proglottids or adult nematodes like Ascaris or
on clinical signs and symptoms alone.
Enterobius may be found on or beneath the
A parasitology laboratory should be able to: surface of the sample.

 Confirm a clinical impression that the

condition has a parasitic nature.
DIRECT FECAL SMEAR
 Rule out differential diagnosis
 Aid a clinician in the choice of proper PRINCIPLES
medication; and
 DIRECT FECAL SMEAR (DFS)- simplest and most
 Help in monitoring the effect of a treatment
rapid of all fecal examination technique.
regimen.
 Recommended for identification of Protozoan
 Diagnosis of parasitic infectious is done either
Trophozoites and detection of Helminthic
by the demonstration of parasite or parasite
Infection.
components (e.g., adult, eggs, larva, cysts,
 One DFS Preparation contains approximately 2mg
oocysts, trophozoites, and antigen) or by the
of feces.
detection of host immune response to the
 Saline and / or Lugol’s Iodine Solution can be
parasite (e.g., antibodies)
used to emulsify the fecal material.
 Demonstration of parasites is possible only
 A. UNSTAINED FILM- useful for the study of living
during the patent stage of the infection
parasite objects. (e.g., Motile Protozoan
 Trophozoites, Helminth Eggs, and Nematodes  It is a QUANTITATIVE METHOD for counting
Larva. Helminth Eggs
 B. IODINE FILM- employed to study the diagnostic  The number of eggs per gram (NPEG) of feces can
features of Protozoan Cysts. be computed by multiplying the number of eggs
observed per thick by 24.

CELLULOSE TAPE PERIANAL SWAB

CONCENTRATION TECHNIQUES
PRINCIPLE
Concentration Techniques can separate Protozoan cysts
 Pinworm Infection (Enterobius Vermicularis) is
and Helminth Eggs from a larger amount of stool (usually
suspected in children with Perianal Itching,
1g in amount) based on differences in specific gravity.
Insomnia, and Restlessness
 Evidence depends on recovery of Adult Worms, 1. SEDIMENTATION PROCEDURE- will sink.
Eggs, or Both a) Acid Ether Concentration Technique
 They are RARELY found in stool examination. (AECT)
 Pinworm Infection can be be best diagnosed by b) Formalin-Ether/Ethyl Acetate
swabbing perianal area using Graham’s Concentration Technique (FECT)
Cellophane (Cellulose) Tape Method 2. FLOTATION PROCEDURE- will float.
a) Zinc Sulfate (ZnSO4) Flotation
GRAHAM” S CELLOPHANE TAPE METHOD b) Brine Flotation
- Highest sensitivity and specificity results c) Sheather’s Sugar Flotation
- Best to collect specimens in the morning before
the patients bathes of defecates.
STOOL CULTURE METHODS

I. Stools positive for hookworm ova and/or

KATO THICK SMEAR (CELLOPHANE THICK SMEAR) Strongyloides Rhabditiform larvae can be
PRINCIPLE cultured until filariform larvae develop. This
technique can be also used for Trichostrongylus
Glycerin-Malachite Green solution Sp.
 GLYCERINE- clears the fecal film to visualize the II. CORPO CULTURE
Helminth Eggs III. HARADA-MORI or THE TEST TUBE CULTURE
 MALACHITE GREEN- dye used to protect the eyes METHOD
from intense light needed in examining the thick
smear.
 Preparation; 500ml distilled water + 500ml STAINING OF STOOL SPECIMEN
Glycerine + 5ml 3% Malachite Green Solution in Staining of stool specimen can also be done specifically in
H2O. the examination of the nuclear characteristics of
NOTE: This method is not suitable for Diarrheic Stool and amoebae.
CAN’T detect Protozoan Cysts and Trophozoites. 1) Iron-Hematoxylin
2) Trichome
3) Periodic Acid Schiff (PAS)
KATO-KATZ THICK SMEAR (MODIFIED THICK SMEAR) 4) Chlorazol Black E
PRINCIPLE

 Kato-Katz thick Smear is a modification of Kato-

Thick Smear Method