Christensen Preben, Knudsen Dorte B, Wachmann Henrik, Madsen Mads T
Section for Reproduction, Department of Large Animal Sciences, Royal Veterinary and Agricultural University, Dyrlaegevej 68, 1870 Frederiksberg C, Denmark.
Theriogenology. 2004 Oct 1;62(7):1218-28. doi: 10.1016/j.theriogenology.2004.01.015.
A simple and rapid flow cytometric method has recently been developed for simultaneous determination of sperm concentration and viability in semen from domestic animals. Use of SYBR-14 trade mark in combination with propidium iodide (PI) allows estimation of the proportion of live sperm (viability). An internal standard of fluorescent microspheres (beads) makes it possible to determine the sperm concentration during the same analysis. In the first experiment, the relationship between sperm viability and litter size was investigated. The second experiment explored whether a smaller variation in the number of motile sperm per insemination dose could be obtained using the FACSCount AF flow cytometer than using a spectrophotometer. Results in the first experiment show that sperm viability is closer related to litter size than is the traditionally used motility parameter. Although the flow cytometer is precise and objective, a limited effect on litter size should be anticipated if ejaculates are selected for insemination according to the percentage of viable sperm. However, the present trial used large insemination doses (2.3 x 10(9) motile sperm/dose) which partially compensate for the differences in motility and viability between boars and ejaculates. In the second experiment it was found that variation in the number of motile sperm per insemination dose could be reduced significantly if the FACSCount AF flow cytometer rather than the Corning 254 spectrophotometer was used for determination of sperm concentration in the raw semen. It is concluded that the FACSCount AF flow cytometer is a strong tool for improvement of the quality control in artificial insemination (AI) centres.
最近开发了一种简单快速的流式细胞术方法，用于同时测定家畜精液中的精子浓度和活力。使用SYBR-14商标与碘化丙啶（PI）结合可估计活精子的比例（活力）。荧光微球（珠子）的内标使得在同一次分析中能够确定精子浓度。在第一个实验中，研究了精子活力与产仔数之间的关系。第二个实验探讨了使用FACSCount AF流式细胞仪是否比使用分光光度计能使每次输精剂量中活动精子数量的变化更小。第一个实验的结果表明，与传统使用的活力参数相比，精子活力与产仔数的关系更为密切。尽管流式细胞仪精确且客观，但如果根据活精子的百分比选择射精用于输精，预计对产仔数的影响有限。然而，本试验使用了大剂量的输精（2.3×10⁹个活动精子/剂量），这部分弥补了公猪和射精之间活力和活力的差异。在第二个实验中发现，如果使用FACSCount AF流式细胞仪而不是康宁254分光光度计来测定原精液中的精子浓度，每次输精剂量中活动精子数量的变化可以显著降低。得出的结论是，FACSCount AF流式细胞仪是改善人工授精（AI）中心质量控制的有力工具。
