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Microbiology and Immunology

Microbiology and Immunology

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Published by: chiprios on Feb 28, 2011
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The saponin lysis method (32) can be performed on either EDT A-
or citrate-anticoagulated blood. Saponin solution to lyse erythrocytes
is available in most laboratories for use with automated hematology

1. Centrifuge up to 10 m1 of blood at 150 x g for 10 min.

2. Remove and discard the plasma.

3. Mix the packed erythrocytes with 50 m1 of 0.5% saponin solution

in 0.85% saline.

4. Mix the solution at intervals for 15 min.
5. Centrifuge the solution at 650 x g for 10 min.
6. Decant and discard the supernatant (there should be about 1 m1

of sediment).

7. Spread several drops of sediment on a glass slide (1 by 3 in.),
and examine two such uncovered wet mounts for motile microfilariae.
Allow the wet mounts to dry before they are fixed and stained.
8. Prepare four or five similal wet mounts and examine them as



described above. To each slide, immediately add 2 drops of 1 % acetic
acid solution and mix it well (microfilariae will be killed and
straightened). Allow the slide to air dry.
9. Dip the dried slides in buffered methylene bluephosphate solution.
10. Rinse the slides in distilled water, and let them air dry.
11. Stain the mounts for 10 min in a 1: 20 dilution of Giemsa
stain in buffered water.
12. Examine the slides microscopically.

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