Hidalgo Manuel, Rodríguez Inmaculada, Dorado Jesús
Reproduction and Obstetrics Unit, Faculty of Veterinary Science, University of Cordoba, 14071 Cordoba, Spain.
Theriogenology. 2006 Sep 1;66(4):996-1003. doi: 10.1016/j.theriogenology.2006.02.039. Epub 2006 Apr 18.
Computer-assisted sperm morphometry analysis (ASMA) has improved the assessment of sperm morphology, but the results depend on the use of adequate sampling and staining procedures of spermatozoa from individual species. In this study, the Sperm Class Analyzer ASMA system was used for the morphometric analysis of goat sperm heads. Semen samples, obtained from four bucks, were used to evaluate the influence of three staining procedures (Diff-Quik, Hemacolor and Harris' Haematoxylin) on the accuracy of image processing and sperm morphometry, the effect of the number of cells analysed and the repeatability of the method. These experiments were performed to obtain objective, accurate and reliable sperm morphometric measurements of goat spermatozoa. Diff-Quik and Harris' Haematoxylin were significantly (p<0.05) more accurate than Hemacolor. However, Diff-Quik obtained the highest proportion of correctly analysed sperm heads (86.06%) and the lowest coefficients of variation on the image processing and morphometric measurements. The staining methods affected significantly the sperm dimensions (p<0.001) with increased values from Diff-Quik than Hemacolor and Harris' Haematoxylin, respectively (Diff-Quik>Hemacolor>Harris' Haematoxylin). No differences in morphometric parameters were found when 100, 150, 175 or 200 spermatozoa were analysed. The repeatability of results obtained was very high since no differences were found when measuring the same sperm on multiple attempts. In conclusion, to obtain objective, accurate and repeatable sperm morphometric measurements by the Sperm Class Analyzer system in goats, the analysis of 100 spermatozoa from slides which have been previously stained with Diff-Quik is recommended.
计算机辅助精子形态测量分析（ASMA）改善了精子形态的评估，但结果取决于对来自各个物种的精子采用适当的采样和染色程序。在本研究中，精子分类分析仪ASMA系统用于山羊精子头部的形态测量分析。从四只公羊采集的精液样本用于评估三种染色程序（Diff-Quik、Hemacolor和Harris苏木精）对图像处理和精子形态测量准确性的影响、分析细胞数量的作用以及该方法的可重复性。进行这些实验是为了获得山羊精子客观、准确和可靠的形态测量数据。Diff-Quik和Harris苏木精的准确性显著高于Hemacolor（p<0.05）。然而，Diff-Quik在正确分析的精子头部比例最高（86.06%），并且在图像处理和形态测量方面的变异系数最低。染色方法对精子尺寸有显著影响（p<0.001），Diff-Quik处理后的精子尺寸值分别高于Hemacolor和Harris苏木精（Diff-Quik>Hemacolor>Harris苏木精）。分析100、150、175或200个精子时，形态测量参数没有差异。多次测量同一精子时未发现差异，因此所获结果的可重复性非常高。总之，为了通过精子分类分析仪系统在山羊中获得客观、准确和可重复的精子形态测量数据，建议对预先用Diff-Quik染色的载玻片上的100个精子进行分析。
