Performing a Manual Count for Sperm Concentration

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Performing a Manual Countfor Sperm Concentration

Dr. Bruce Eilts Louisiana State University
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Diluent Formol Buffered Saline (Water works
too!)
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A one page how to. Proceed for step by step
instructions.
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You have now created a 1100 dilution and are
ready to load the hemacytometer
Click once to proceed!
Raw Semen
FBS or H2O
0.1 mL?1100
0.1 mL
0.1 mL
0.9 mL
0.9 mL
0.9 mL
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Neubauer chamber
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Load each side of the chamber.
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Load each side of the chamber.
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(No Transcript)
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40x Objective Phase ideal Match 40x phase ring
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Phase contrast makes counting much easier. If you
do not have a phase contrast microscope, you will
have to decrease the light by closing the
diaphragm.
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There are 9 big squares on the hemacytomter
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1 2 3 4 5 6 7
8 9
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Count the middle big square bounded by triple
lines.
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There are 25 smaller squares within the middle
big square bounded by triple lines.
1 2 3 4 5 6 7 8 9 10 11
12 13 14 15 16 17 18 19 20 21 22 23
24 25
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Each of the 25 smaller squares has 16 smaller
squares
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Count each cell in the 25 squares
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Count cells on the left and top lines or on the
right and bottom lines as you go to prevent from
counting cells twice
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How many cells do you count here?
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You should get 32 Click to follow the counting
ball.
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Count both sides of the chamber. Results should
be within 10 of each other. If the results are
not within 10 of each other, repeat the entire
counting procedure.
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• Number of cells counted in the 25 squares
• Sperm X 106/mL

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• Number of cells counted in the 25 squares 32 in
  our example.
• Sperm concentration
• 32 X 106/sperm cells/mL

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• If sperm concentration
• 32 X 106/sperm cells/mL
• and
• Volume 15 mL
• Total count is
• 480 X 106

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• You have finished successfully!