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Overview :
Use the hemacytometer to get a count of spermatozoa per mL
semen.
Diluent:
(100 mL distilled water) + (1 mL neutral buffered formalin) +
(5 grams sodium bicarbonate)
Procedure:
1) Make a
1:20 dilution with 0.5 mL of semen diluted to 10 mL. Mix.
2) Charge
both sides of the hemacytometer and wait 2 minutes for the immobilized sperm
(fixed by formalin) to settle.
3) Count
the central chamber (square #5) on both sides of a hemocytometer and add the
numbers together. Each square is 1 mm square with a depth of 0.1 mm, for a
volume of 0.1 cumm. According to convention, elements are counted if they lie on
the topmost and leftmost lines, but not the bottommost or rightmost lines.
4) Repeat
the count on a fresh sample.
5) Average
the two counts.
6) Multiply
the average by 100,000, to get total spermatozoa per mL.
where:
• 100,000
(from step 6) = ((dilution factor 20) / ((number of squares 2) * (volume 0.1
cumm))) * (1000 cumm/mL)]
Limitations:
• The test shows considerable intra- and inter-technologist
variation.
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