spermatozoa staining

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
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For ram semen testing,

A.  2 gm eosin in 100 ml M/8 phosphate buffer, pH 7.2

B.  1.5% aniline blue in absolute ethanol aniline blue has a solubility of
approx 1.1% in alcohol, must be a sat solution

one drop stain on slide, one drop semen, mix.

Lay second slide on top, and draw them apart like you are doing a bone
marrow smear, to make a thin film, dry on a warm plate at 40C, examine with
oil immersion.

Rose Bengal stain

Sperm smear on slide, the fix in 95% ethanol for 15 min

Air dry slide before staining.

0.25% crystal violet 2 -3 min

distilled water rins

decolorize with 95% ethanol for 20 seconds

wash in water,

1% rose bengal, 1 1/2 min (recommended 10 - 30) better staining longer

wash in water, air dry and mount with Permount

head light color, tail dark color

Came from my old Med TEch text book, Todd and Sanford, 13th edition, p 926,
published in the Dark Ages.

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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