STEP 3: Tap the slide on paper towels. Immerse the slide into solution B and dip two
to three times.
STEP 4: Tap the slide on paper towels. Immerse the slide into distilled water and dip
two to three times.
STEP 5: Place the slide vertically on a slide rack and allow the slide to air dry.
(3) Staining reaction. The malaria parasites are stained similar to staining
with Giemsa (the red chromatin dots with light blue cytoplasm). However, there is
usually substantial debris in the background. Thus, it is an excellent stain for mass
production in an epidemic situation. Its use is limited to thick drop preparations. It is not
recommended for thin films.
d. Jaswant-Singh-Bhattacherji (JSB) Stain. The JSB stain is a fairly rapid
staining method for the detection of malarial parasites. This stain is superior to the
Field's stain because the parasites stain clearer and both thick and thin smears can be
stained. However, preparations fade quite rapidly. Therefore, this stain is not
recommended when permanent slides are desired.
(1)
Preparation of reagents.
(a) Solution A.
Medicinal Methylene blue.................................. 0.5 gm
Potassium dichromate....................................... 0.5 gm
Sulfuric acid (1.0 percent solution) .................... 3.0 ml
Potassium hydroxide (1.0 percent solution) ...... 10.0 ml
Distilled water .................................................... 500.0 ml
STEP 1: Dissolve the methylene blue in the 500 milliliters of distilled water.
STEP 2: Add the sulfuric acid gradually while stirring. After mixing the solution well,
add the potassium dichromate. A heavy amorphous purple precipitate of
methylene blue/dichromate forms immediately.
STEP 3: Place the container with the precipitate in an autoclave for three hours at a
temperature of 100-109 C and five pounds pressure until the purple color
shifts to a deep blue. Allow the product to cool to room temperature. Add the
potassium hydroxide slowly, drop by drop, while stirring.
STEP 4: Mix the solution well and age at room temperature for 48 hours. Filter and
store in an amber-colored bottle.
MD0841
2-20