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COMPARISON OF THREE METHODS FOR COUNTING HUMAN SPERMATOZOA
Citation:
Jeffay, S C., L F. Strader, R A. Morris, J E. Schmid, Olshan. AF, L. W. Lansdell, AND S P. Darney. COMPARISON OF THREE METHODS FOR COUNTING HUMAN SPERMATOZOA. Presented at American Society of Andrology, Phoenix, AZ, March 26-April 2, 2003.
Description:
COMPARISON OF THREE METHODS FOR COUNTING HUMAN SPERMATOZOA SC Jeffay1, LF Strader1, RA Morris1, JE Schmid1, AF Olshan2, LW Lansdell2, SD Perreault1. 1US EPA/ORD, RTP, NC; 2UNC-CH, Chapel Hill, NC.
The IDENT feature of the HTM-IVOS semen analyzer (Hamilton Thorne Research, Beverly, MA) allows rapid automated sperm counts to be obtained from multiple fields. In an ongoing epidemiology study we are comparing IDENT sperm counts with those obtained manually using the same microscope chamber (12mm deep microcell slide) or with a hemacytometer. 100ml semen (n=36 samples) was added to 100ml Hoechst IDENT stain after which the stained sperm suspension was further diluted as needed to obtain optimal concentrations for each chamber. Diluted semen was loaded into the microcell and 20 fields were counted on each side by IDENT. The same chamber was transferred to a microscope and 5 fields counted on each side at 500x using an eye piece reticle (MC). Diluted semen was also loaded into a hemacytometer (H) and both sides counted at 200x. Sperm counts (millions/ml) were log transformed and analyzed by ANOVA (SAS, Cary, NC). After adjusting for sample ID, the mean log count (? sem) was significantly higher for H than for IDENT or MC (least squares means: 1.97?0.01*, 1.86?0.01, 1.87?0.01, respectively, p<0.001). The variance due to chamber side (replicate) after removing that for sample ID, was lower for IDENT than MC or H (mean square error: 0.0008, 0.003 and 0.002, respectively). The absolute difference between replicates was also lowest for IDENT: least squares means were IDENT: 0.030 ? 0.006*, MC: 0.069 ? 0.006, and H: 0.53 ? 0.006 (p<0.01). We conclude that 1) IDENT and manual sperm counts performed in the MC chamber are similar and lower than H, and 2) IDENT provides a more repeatable and efficient method for counting human sperm than MC or H. Supported by CR829327; this abstract does not reflect EPA policy.