Validation of a comercially available fluorescence-based instrument to evaluate stallion spermatozoal concentration and comparison to photometric systems

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Title: Validation of a comercially available fluorescence-based instrument to evaluate stallion spermatozoal concentration and comparison to photometric systems
Author: Comerford, Kathryn L.
Abstract: Accurate measurement of stallion spermatozoal concentration is important to equine breeding operations . The hemacytometer is considered the standard for measuring spermatozoal concentration but is time consuming and may be imprecise . The flow cytometer is considered precise and accurate , but only practical for research purposes due to sample preparation time and high cost . Photometric systems are commonly used but can be inaccurate outside a relatively narrow concentration range and can be rendered inaccurate in the presence of contaminants . A new instrument , the NucleoCounter SP -100 is reported to enumerate spermatozoa at wider concentration ranges and can identify spermatozoa in opaque semen extenders . Epididymal , neat (raw ) ejaculates , and ejaculates diluted in various semen extenders were analyzed with the NucleoCounter , the Densimeter , the Spermacue , flow cytometric and hemacytometric methods . Results were compared statistically by : 1 ) regression analysis , 2 ) the agreement of two instruments , whereby the difference in values between two instruments was plotted on the y -axis against the mean of those values on the x -axis [26] and 3 ) a modified method that measured the percentage deviation , whereby the percentage (of the difference in values between two instruments divided by the mean ) of the same two values was plotted on the y -axis against the mean value of the two instruments on the x -axis . The NucleoCounter showed more agreement with both the flow cytometer and hemacytometer for epididymal , neat ejaculated and extended spermatozoa over a range of concentrations than the Densimeter or the Spermacue . The NucleoCounter showed more agreement with the flow cytometer for epididymal and neat ejaculated spermatozoa and more agreement with the hemacytometer for spermatozoa diluted in semen extenders . The Spermacue showed the least agreement with both standards for all spermatozoal comparisons . All coefficients of variation for the flow cytometer , hemacytometer and NucleoCounter were >10 % for all spermatozoal comparisons . This study indicates that the NucleoCounter shows more agreement with the flow cytometer and hemacytometer than photometric systems when evaluated with epididymal , neat ejaculated and extended spermatozoa . The instrument is also more repeatable than either photometric system , but may be cost -prohibitive for some operations .
URI: http : / /hdl .handle .net /1969 .1 /ETD -TAMU -2009 -05 -261
Date: 2010-01-16

Citation

Validation of a comercially available fluorescence-based instrument to evaluate stallion spermatozoal concentration and comparison to photometric systems. Available electronically from http : / /hdl .handle .net /1969 .1 /ETD -TAMU -2009 -05 -261 .

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