A technical note on diluting semen for the haemocytometric determination of sperm concentration

A comparison was made of the use of either an SMI positive displacement pipette or an Eppendorf Varipette as the method of sampling 63 liquefied semen samples for volumetric dilution and haemocytometry to determine sperm concentration. The 95% range of the differences between the values obtained using the SMI and the Varipette with whole tips was from -46.9 to 63.8 x 10(6)/ml. With the Varipette tips cut off 12.5 mm from the end the 95% range was from -52.4 to 55.8 x 10(6)/ml. Previous work had shown that the 95% ranges of differences between duplicate determinations using the SMI pipette were from -7.2 to 6.9 x 10(6)/ml for two 1 + 19 dilutions, or from -16.0 to 12.1 x 10(6)/ml for 1 + 19 and 1 + 49 parallel dilutions. Therefore, since the Varipette had a far greater potential for error, a positive displacement pipette should be used when taking precise volume aliquots of human semen.