Farrell P B, Foote R H, McArdle M M, Trouern-Trend V L, Tardif A L
Department of Animal Science, Cornell University, Ithaca, New York 14853-4801, USA.
J Androl. 1996 May-Jun;17(3):293-300.
Proper handling of semen prior to computer-assisted sperm analysis (CASA) is critical if the analysis is to be representative of the fresh sample. The effects of diluting medium or dilution and holding time before CASA on multiple sperm characteristics were studied. Four replicates of unselected semen samples from each of eight human donors were diluted with phosphate-buffered saline (PBS)-glucose plus bovine serum albumin (BSA), with Tyrode's albumen lactate pyruvate (TALP), and with high-potassium TALP (K-TALP) to a concentration of approximately 25 x 10(6) sperm/ml. The diluted semen was held for 0, 1, and 2 hours at approximately 30 degrees C before CASA, with little difference between the three diluents in all 12 variables measured. There was a decline of 3-6% in the proportion of motile sperm over a 2-hour period (P < 0.05). Donors were the largest source of differences (P < 0.05). Rabbit sperm (five bucks, four ejaculates per buck) were processed in a manner similar to that of the human sperm. There was a major effect of media. The average percentages of motile sperm over 2 hours in TALP, K-TALP, and PBS were 76, 42, and 29%, respectively (P < 0.05), with a decline of only 3% in TALP during the 2 hours. Hyperactivity and other characteristics were affected by treatment. Donors were a large source of variation. Bull semen (10 bulls, two ejaculates per bull) either was not diluted or diluted with TALP 2x or 4x and held for 0, 1, and 2 hours at 30 degrees C. It was then diluted to 25 x 10(6) sperm/ml with TALP. There was little change in most sperm characteristics in any treatment during the first hour, although many of the changes were statistically significant. The percentage of motile sperm in undiluted semen declined from 87% to 82% over 2 hours. Modified TALP was a suitable medium for sperm from all three species, and a simple PBS-glucose-BSA medium can be used for human sperm.
如果计算机辅助精子分析（CASA）要代表新鲜样本，那么在分析前对精液进行适当处理至关重要。研究了CASA前稀释介质、稀释及保存时间对多种精子特征的影响。从8名人类供体采集的未筛选精液样本各取4份，分别用磷酸盐缓冲盐水（PBS）-葡萄糖加牛血清白蛋白（BSA）、台氏白蛋白乳酸丙酮酸（TALP）和高钾TALP（K-TALP）稀释至浓度约为25×10⁶精子/ml。在CASA前，将稀释后的精液在约30℃下保存0、1和2小时，在所有测量的12个变量中，三种稀释剂之间差异不大。在2小时内，活动精子比例下降了3%-6%（P<0.05）。供体是差异的最大来源（P<0.05）。兔精子（5只雄兔，每只雄兔4次射精）的处理方式与人类精子类似。介质有主要影响。在TALP、K-TALP和PBS中，2小时内活动精子的平均百分比分别为76%、42%和29%（P<0.05），在TALP中2小时内仅下降了3%。超活性和其他特征受处理影响。供体是变异的一个重要来源。公牛精液（10头公牛，每头公牛2次射精）要么不稀释，要么用TALP 2倍或4倍稀释，并在30℃下保存0、1和2小时。然后用TALP将其稀释至25×10⁶精子/ml。在第一个小时内，任何处理中大多数精子特征变化不大，尽管许多变化具有统计学意义。未稀释精液中活动精子的百分比在2小时内从87%降至82%。改良TALP是适合所有三种物种精子的介质，简单的PBS-葡萄糖-BSA介质可用于人类精子。
