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Delay in the application of the cover glass is a potential source of error with the Makler Counting Chamber
1Concept Fertility Centre, King Edward Memorial Hospital, Subiaco, Western Australia, Australia. concept@conceptfert.com.au
Objective:
To determine the effect of delaying the application of the cover glass of a Makler Counting Chamber (Sefi Medical Instruments, Haifa, Israel) on the apparent concentration of sperm or Dynabeads (Dynal Pty Ltd, Carlton, Victoria, Australia).
Design:
Systematic alteration of the time taken to apply the cover glass.
Setting:
Comprehensive infertility service.
Patient(S):
Fresh semen from a patient or washed sperm from a donor.
Intervention(S):
For the Makler Counting Chamber, a 5-microL drop of the sperm or bead suspension was placed in the center of the lower platform and the cover glass was applied either immediately (0 seconds) or after a delay of 5, 10, or 30 seconds.
Main Outcome Measure(S):
The concentration of beads or sperm counted using an Improved Neubauer Hemocytometer or a Makler Counting Chamber.
Result(S):
There was a progressive increase in sperm concentration with a longer delay in applying the cover glass when sperm was suspended in either seminal plasma or culture medium. Repeating the experiments with Dynabeads resuspended in phosphate-buffered saline, serum, or seminal plasma showed the same trend, although the suspension of beads in serum seemed to settle the least over the first 10 seconds.
Conclusion(S):
Delay in the application of the cover glass is a potential source of error with the Makler Counting Chamber and should be avoided.
