Preservative Advantages Disadvantages

10% Formalin
All-purpose fixative Not suitable for some permanent
Neutralized formalin Easy to prepare smears stained with trichrome
Procedure
Long shelf life Inadequate preservation of
- Neutralize the formic acid (in the formalin) by adding
Good preservation of morphology of helminth eggs, morphology of protozoan trophozoites
10 N NaOH until the pH becomes neutral (check with pH
paper or on a pH meter). larvae, protozoan cysts, and coccidia Can interfere with PCR, especially

- To fix stool specimens, a 10% neutralized formalin Suitable for concentration procedures and UV after extended fixation time
solution is generally used. To fix protozoa, a 5% solution fluorescence microscopy
is recommended.
Suitable for acid-fast, safranin, and chromotrope
Neutralized formalin solution (10%)
stains
Procedure Compatible with immunoassay kits and UV
- Combine 100 ml neutralized formalin with 900 ml fluorescence microscopy
distilled water or saline;

- Mix well. Store in a sealed container.

Neutralized formalin solution (5%)

Procedure

- Combine 50 ml neutralized formalin with 950 ml

distilled water or saline;

- Mix well. Store in a sealed container.

MIF
merthiolate-iodine-formaldehyde) Components both fix and stain organisms Not suitable for some permanent
Easy to prepare smears stained with trichrome
MIF solution for staining fresh stool specimens
Composition Long shelf life Inadequate preservation of
Useful for field surveys morphology of protozoan trophozoites
- Merthiolate tincture, 1/1000 Lilly 7.75 ml
Suitable for concentration procedures Iodine interferes with other stains and
- Lugol's 5% iodine solution 1.00 fluorescence
ml
Iodine may cause distortion of
- Formalin 37%-40% 1.25
protozoa
ml

LV-PVA
(low viscosity polyvinyl-alcohol) Good preservation of morphology of protozoan Inadequate preservation of
trophozoites and cysts morphology of helminth eggs and
Composition
Easy preparation of permanent smears stained with larvae, coccidia, and microsporidia
- Schaudinn's stock solution 93.5 ml
such as trichrome (solution both preserves Contains mercuric chloride
- Glycerol 1.5 ml organisms and makes them adhere to slides) Difficult and expensive to dispose of

- Glacial acetic acid 5.0 ml Preserved samples remain stable for several months Difficult to prepare in the laboratory
Not suitable for concentration
- Polyvinyl alcohol 90-50 (low or medium viscosity)
5g procedures

Procedure Cannot be used with immunoassay

kits
- Combine the first three ingredients and heat the
mixture in a water bath (75 C), making sure that the Not suitable for acid-fast, safranin and
ethanol (in Schaudinn's solution) does not burn
chromotrope stains
- Slowly add 5 g polyvinyl alcohol, stirring continuously;
if possible, use a heating plate with a magnetic stirrer.

SAF
Suitable for both concentration procedures and Requires additive (e.g., albumin-
 (sodium acetate-acetic acid-formalin)
preparation of permanent stained smears glycerin) for adhesion of specimens to
Composition Easy to prepare slides

- Anhydrous sodium acetate (NaC2H3O2) 9g Long shelf life Permanent stains not as good as with
Suitable for acid-fast, safranin, and chromotrope PVA or Schaudinn's fixative
- Glacial acetic acid 20 ml
stains
- Commercially available formalin (37%-40%) 40 ml
Compatible with immunoassay kits
- Distilled water 925 ml

In alternative to 9 g anhydrous sodium acetate, 15 g

sodium acetate trihydrate (CH3CO2Na 3H2O) can be
used.

Procedure

- Dissolve the salt in distilled water;

- Add formalin and glacial acetic acid, and mix well;

- Adjust to pH 4.15, if necessary.

Schaudinns Fixative
Schaudinn's stock solution Good preservation of morphology of protozoan Less suitable for concentration
Composition trophozoites and cysts procedures

- Saturated mercuric chloride solution 2 volumes Easy preparation of permanent stained smears Contains mercuric chloride
Inadequate preservation of
- 95% ethanol 1 volume
morphology of helminth eggs and
- Mix well.
larvae, coccidia, and microsporidia
Schaudinns working solution Poor adhesion of liquid or mucoid
- Immediately before use, add 5.0 ml glacial acetic
acid to 95 ml of stock solution specimens to slides

- Mix well