STOOL

PRESERVATION
LEARNING OUTCOMES
● At the end of this lesson, the student is expected to:
○ Know the definition and purpose of stool fixatives
○ Identify indications for the use of stool fixatives
○ Determine the factors to consider in collection, processing and
examination prior to fixation
○ Enumerate the commonly used
 WHAT IS A STOOL FIXATIVE?

● FIXATIVES
○ Preserve the morphology of protozoa
○ Prevent further development of certain helminth eggs and larvae
● Available commercially or prepared in the lab
● Recommended ratio = 3:1 (fixative : stool)
 WHY PRESERVE SPECIMENS?

● INDICATIONS:

○ Fresh stool samples are not possible

○ Specimen examination delay

○ Collecting specimens for many days

○ Preparing permanent stains

● Prior to processing:

○ Fix specimens for at least 30 minutes

 COMMONLY USED FIXATIVES

● FORMALIN

• 5%- ideally preserves protozoan cysts

• 10% - preserves helminth eggs and larvae

● MERTHIOLATE IODINE FORMALDEHYDE (MIF)
● LOW-VISCOSITY POLYVINYL ALCOHOL (LV-PVA)
● SHAUDINNS’ FIXATIVE
● SODIUM ACETATE ACETIC ACID-FORMALIN (SAF)
● MODIFIED PVA
● ONE-VIAL FIXATIVES
SPECIMEN CONSIDERATIONS
 FORMALIN

ADVANTAGES DISADVANTAGES
1. All purpose fixatives 1. Not suitable for some permanent
2. Easy to prepare smears with trichrome
3. Long shelf life 2. Can interfere with PCR, especially
4. Good preservation of helminth after extended fixation time
eggs, larvae, protozoan cyst, and 3. Inadequate preservation of
coccidia morphology of protozoan
5. Suitable for concentration trophozoites.
procedures
6. Suitable for acid-fast, safranin, and
chromotrope stains
7. Compatible with immunoassay kits
 MERTHIOLATE IODINE FORMALDEHYDE
(MIF)
ADVANTAGES DISADVANTAGES
1. Components both fix and stain 1. Not suitable for some permanent
organisms smears with trichrome
2. Easy to prepare 2. Inadequate preservation of
3. Long shelf life morphology of protozoan
4. Suitable for concentration trophozoites
procedures 3. Iodine interferes with other stains
5. Useful for field surveys and flourescense
4. Iodine may cause distortion of
protozoa
 LOW-VISCOSITY POLYVINYL ALCOHOL
(LV-PVA)
ADVANTAGES DISADVANTAGES
1. Preserves and allows organisms to 1. Inadequate preservation of
adhere to slides well morphology of helminth egg and
-adhesive for the stool specimen when larvae, coccidia, and microsporidia
preparing slides for staining 2. Contains mercuric chloride
2. Preserved samples remain stable for 3. Difficult and expensive to dispose
several months 4. Difficult to prepare in the laboratory
3. Good preservation of morphology of 5. Not suitable for concentration
protozoan trophozoites and cysts procedures
4. Easy preparation of permanent 6. Cannot be used with immunoassay
smears stained such as trichrome kits
7. Not suitable for acid-fast, safranin
and chromotrope stains
 SHAUDINN’S FIXATIVE

ADVANTAGES DISADVANTAGES
1. Good preservation of morphology 1. Contains mercuric chloride
of protozoan trophozoites and cysts 2. Inadequate preservation of
2. Easy preparation of permanent morphology of helminth eggs and
stained smear larvae, coccidia, and microsporidia
3. Less suitable for concentration
procedures
4. Poor adhesion of liquid or mucoid
specimens to slides
 SODIUM ACETATE – ACETIC ACID FORMALIN
-can be used for performing concentration techniques and permanent stained
smears
ADVANTAGES DISADVANTAGES
1. Easy to prepare 1. Requires additive (e.g. albumin-
2. Long shelf life glycerin) for adhesion of specimens
3. Suitable for concentration to slides
procedures 2. Permanent stains not as good as
4. Suitable for acid-fast safranin, and with PVA or Schaudinn's fixative
chromotrope stains
5. Compatibly with immunoassay
kits
6. Suitable for the preparation of
permanent stained smears
 MODIFIED PVA
-Other alternatives to mercury-based PVA are the use of substitute
compounds containing copper sulfate or zinc sulfate

ADVANTAGES DISADVANTAGES
1. No mercuric chloride 1. Staining not consistent
2. Zinc is preferred over copper 2. Organism morphology maybe poor
3. Permanent smears can be made and 3. Copper-morphology of cysts and
stained with trichrome trophozoites is poor
4. Zinc better morphology but not
comparable to LV-PVA
 ONE-VIAL FIXATIVES
-free of formalin and mercury and can be used for concentration techniques
and permanent stained smears
ADVANTAGES DISADVANTAGES
1. No mercuric chloride 1. Certain one-vial fixatives must use
2. Concentrate and permanent smears certain stains
can be made out of one vial 2. Color difference of stain
3. Immunoassays can be done on most 3. Staining not always consistent
4. Sometimes more expensive than
formalin and LV-PVA
NOTES TO REMEMBER
● Preservation of specimens in necessary when stool sample cannot be
examined within the prescribed time interval
● The choice of preservative will depend on the type of test or stain the
sample will undergo
● The choice of preservative also depends on the laboratory’s
preferences
● The medical technologist should know the advantages and
disadvantages of each commonly used preservative in the lab
● Properly prepare specimens for reservation
Scotch Tape
Method
Vanessa F. Ferriol, RMT
Learning Outcomes

At the end of this lesson, the student is able to:

• Demonstrate the preparation and use of a cellophane tape swab for
perianal specimens;
• Perform the technique skillfully;
• Explain the rationale of doing this procedure for the recovery of
helminth eggs;
• and Appreciate the importance of tracing the life cycle of parasites to
identify their portal of exit and thus perform the accurate laboratory
procedure to demonstrate its diagnostic stage
 Enterobius
vermicularis
• Asymptomatic
• Pruritus ani Insomnia
• Anorexia
• Vulvovaginitis
• Lifecycle: Simple/Direct
Retroinfection Autoinfection
Epidemiology
PRINCIPLE OF THE TEST
● NAMES OF THE TEST

○ CELLOPHANE TAPE SWAB / CELLULOSE TAPE SWAB / SCOTCH TAPE SWAB

● The most widely used procedure for the detection of human pinworm infections
● Timing of spx collection: early morning, before bath or toilet use

● Expected results:

○ Eggs, 50-60 um x 20-30 um

○ Female adult worm

● REPORTING OF RESULTS
○ Enterobius vermicularis eggs present
○ Enterobius vermicularis adult present
○ No Enterobius vermicularis eggs or adult seen
PRINCIPLE OF THE TEST
● Asymmetrical egg with translucent shell of layers

○ Outer triple albuminous layer (mechanical)

○ Inner embryonic lipoidal membrane (chemical)

● Infective: 4-6 hours
● Viability: 13 hours under moist conditions
● MOT: ingestion or inhalation
● Diagnosis: identifying the worm or its eggs
 PROCEDURE
● Fold together sticky surfaces of a piece of clear
cellophane tape
● Stretch tape, sticky side out, over on a wooden tongue
depressor, holding non-sticky ends firmly with thumb
or forefinger
● Apply tape to anal area, rocking back and forth to
cover as much as the mucosa and mucocutaneous area
as possible
● Remove tape and apply tape to microscopic slide,
sticky side down. Press firmly into position.
● Examine for eggs under LPO, then shift to HPO when
E. vermicularis eggs are suspected.
NOTES
● CTS is a very simple, quick, non-invasive, cost-effective procedure used to recover
Enterobius vermicularis ova.

● SPECIMEN CONSIDERATION

○ Use clear cellophane. In case frosted tape was used, a few drops of xylene under
the edge clears the tape so that eggs can be seen

○ To increase chance of finding eggs: sample collection done in 3 separate

mornings

○ Before declaring negative result: up to 6-7 successive morning samples should

be collected.