Calculation of sperm suspension volume to use for fertilizing in 100μl drops
The dilution is 1:2 as 25 µl of sperm suspension has been mixed with 25 µl of cold water
And the number of sperm counted in 100 squares must be divided by 10 (in Makler chamber), and multiplied by 2, to give the actual sperm concentration x106/ml
EXAMPLE
You want a final concentration of 2.0x106 sperm/ml for fertilization so you calculate:
The IVF well already contains 80 µl of fertilization medium + 10 µl IVF medium when the oocytes were transferred, so, addition of approximately 10 µl of sperm suspension would give a total volume of 100 µl
Calculation of sperm suspension volume to use for fertilizing in 500 μl wells
The dilution is 1:2 as 25 µl of sperm suspension has been mixed with 25 µl of cold water
The number of sperm counted in 100 squares must be divided by 10 (in Makler chamber), and multiplied by 2, to give the actual sperm concentration x106/ml
EXAMPLE
You want a final concentration of 2.0x106 sperm/ml for fertilization so you calculate:
The IVF well already contains 400 µl of fertilization medium + 60 µl IVF medium when the oocytes were transferred, so addition of approximately 40 µl of sperm suspension would give a total volume of 500 µl
