Name: Klenn Mae P.
Ortreza Date:__________________ Score:________________
Module 2
PRESERVATION OF SPECIMENS
Objectives
At the end of the activity, the students are expected to:
1. State the purpose for using preservatives.
2. Compare and contrast the different preservatives used in the laboratory.
3. Properly preserve a fecal specimen and prepare slides for a permanent stained smear.
Introduction
Trophozoites of intestinal amoebae die and disintegrate rapidly so it is important to achieve
adequate and proper preservation when specimens cannot be examined immediately (within 30 minutes
of passage). Several preservatives are available. The two general methods include physical and chemical
means. The most popular preservative used by clinical laboratories is Polyvinyl Alcohol-Fixative (PVA
fixative). The purpose of fixatives is to prevent deterioration of fragile forms of parasites. Many
commercially prepared kits contain a 2-vial method for patients to submit stool specimens. One vial of
the kit contains PVA-fixative and the other contains 5 or 10 percent formalin. It is very important that the
proper ratio of fecal specimen and fixative are used. If the patient collects the specimen at home they
should be told to fix the specimens immediately after passage. Some organisms rapidly lose their
characteristic morphology.
Materials and Reagents:
Polyvinyl alcohol fixative Sodium Acetate Formalin fixative Formalin glacial acetic
acid
Glass slides distilled water
Coverslips Vials
Procedures
1. On microscope slides:
A drop of dysenteric stool is placed on a microscope slide and mixed with three drops of
PVA-fixative. The mixture is then smeared over approximately two-thirds of the glass surface and
allowed to dry thoroughly (preferably overnight at room temperature). Dried smears remain
satisfactory for extended periods.
2. In Vials:
A quantity of specimens is thoroughly mixed in a vial containing three parts of PVA
fixative. Smears for staining can be prepared immediately by spreading a drop or two of the
mixture over the surface of a slide. It is important not to have the smears too thick and to allow
them to dry thoroughly.
Questions for Research:
1. What are the advantages and disadvantages of using the different preservatives?
Formalin PVA Sodium Acetate
Advantages ● Fixative for all ● The morphology ● Both
purposes of protozoan concentration
● Helminth eggs, trophozoites and procedures
larvae, cysts has been and the
protozoan cysts, preserved well. preparation of
and coccidia ● Permanent permanent
have good smears stained stained
morphology with trichrome smears are
preservation. are simple to possible with
● Acid-fast, manufacture. this tool.
safranin, and ● For several ● It's simple to
chromotrope months, prepare and
stains are all preserved has a long
compatible. samples are shelf life.
stable. ● Acid-fast,
safranin, and
chromotrope
stains are all
compatible.
Disadvantage ● Some persistent ● Lack of ● Additive is
smears tinted morphological required for
with trichrome preservation of specimen
are not helminth eggs adherence on
acceptable. and larvae, slides.
● Inadequate coccidia, and ● Permanent
preservation of microsporidia. stains aren't
protozoan ● Mercuric chloride quite as good
trophozoites' is present. as PVA.
morphology Disposal is
● Interferes with difficult and
PCR, particularly costly.
after a long ● Concentration
fixation time. methods are not
recommended.
Acid-fast,
safranin, and
chromotrope
stains are not
recommended.
2. What preventive measure do you practice when using the different chemical preservatives?
➢ Wear the required equipment and acquaint yourself with the technique.
➢ Glassware that has been chipped or fractured should not be used.
➢ Always have a second person nearby when working with hazardous items.
➢ Know what to do in an emergency.
➢ Maintain a clean and orderly laboratory.
➢ Use a fume hood for hazardous chemicals and a biosafety cabinet for biohazardous
products (BSC)
➢ Whenever possible, decontaminate.
➢ All processes should be carried out with the goal of reducing the amount of aerosol
produced.
3. In what conditions are preservatives allowable to use?
➢ When stool specimens cannot be inspected within the time frame specified, specimen
preservation is required.
