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Measuring Spore Concentration with Hemacytometer

1. Mix spore solution well.


2. Add 10 l of spore solution to each side of the hemacytometer.
3. Count number of spores in zones A, B, C, D and E on both sides
of the hemacytometer, record them, and calculate the average of
the two sides.
a. If a spore falls on the left or bottom line DO NOT count it.
b. If a spore falls on the right or top line DO count it
A

B
E

4. To determine the number of spores per mL


a. (A + B + C + D + E) x 2000
5. Dilute the spore solution to desired concentration (spores/mL)
a. Use C1V1= C2V2 formula
i. Ex. If spore solution is found to be 67,000 spores/mL
and you want 40mL of 50,000 spores/mL
ii. 67,000 x = 40 * 50,000
a. x = 2,000,000 = 29.85 ~=30mL
b. 67,000
iii. So, you will add 30mL spore solution to 10mL sterile
dH20 (40-30=10) to give a final concentration of
50,000 spores/mL in 40mL
6. You can use the Excel worksheet:
C:\Scab data\Protocols\Current\Hemacytometer Worksheet.xls
to print out a data entry form or to calculate spore concentrations.

Nicki Mundell
Page 1
3/23/2015
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