Mack S O, Wolf D P, Tash J S
University of Texas Medical School, Department of Obstetrics, Gynecology and Reproductive Sciences, Houston 77030.
Biol Reprod. 1988 Mar;38(2):270-81. doi: 10.1095/biolreprod38.2.270.
The CellSoft computer-assisted digital image analysis system was validated for quantitating specific motility parameters in large numbers of human sperm. Motility patterns ranging from linear head trajectories (Type 1) to nonlinear, asymmetric patterns with overlapping trajectory (Type 5) were subjectively identified in semen and washed samples prepared for in vitro fertilization. A representative of each type was used for optimizing the digital imaging set-up parameters, tracking rate, and frequency. Each cell type was also characterized according to the following motility parameters: curvilinear velocity (Vcl), straight line velocity (Vsl), linearity of forward progression (Lin), maximum and mean lateral head amplitude (maxLHA; mean LHA), and beat cross frequency (BCF). Comparison of all parameters that could be determined both digitally and manually (Vcl, Vsl, Lin, and BCF) indicated no differences (p greater than 0.05) in Vcl, Lin, or BCF and only slight differences (5-6%) in Vsl measurements. After validation of the digital imaging technique, populations of seminal and washed cells were studied. Replicate analysis of the same sample demonstrated no significant intraassay variability. A comparison of semen and washed cells from 10 different donors indicated that all of the motility parameters, with the exception of Lin, were significantly higher (p less than 0.05) in washed cells. It was concluded that the digital imaging system can adequately and rapidly quantitate a large number of cells with heterogeneous motility patterns. This technique may prove to be useful in defining motility characteristics associated with capacitation, the acrosome reaction, and fertility of human sperm.
CellSoft计算机辅助数字图像分析系统已通过验证，可用于定量大量人类精子的特定运动参数。在为体外受精准备的精液和洗涤样本中，主观识别出了从线性头部轨迹（1型）到非线性、不对称且轨迹重叠的模式（5型）等多种运动模式。每种类型的一个代表样本用于优化数字成像设置参数、跟踪速率和频率。每种细胞类型还根据以下运动参数进行了表征：曲线速度（Vcl）、直线速度（Vsl）、向前运动的线性度（Lin）、最大和平均头部横向振幅（maxLHA；mean LHA）以及鞭打交叉频率（BCF）。对所有可通过数字和手动方式测定的参数（Vcl、Vsl、Lin和BCF）进行比较表明，Vcl、Lin或BCF没有差异（p大于0.05），Vsl测量中只有轻微差异（5 - 6%）。在数字成像技术验证后，对精液细胞群体和洗涤后的细胞群体进行了研究。对同一样本的重复分析表明，测定内没有显著的变异性。对来自10个不同供体的精液和洗涤细胞进行比较表明，除Lin外，所有运动参数在洗涤后的细胞中均显著更高（p小于0.05）。得出的结论是，数字成像系统能够充分且快速地定量大量具有异质运动模式的细胞。该技术可能被证明在定义与人类精子获能、顶体反应和生育能力相关的运动特征方面是有用的。
