Takács T, Szöllösi J, Balázs M, Gáspár R, Mátyus L, Szabó G, Trón L, Resli I, Damjanovich S
Department of Biophysics, University Medical School of Debrecen, Hungary.
Acta Biochim Biophys Hung. 1987;22(1):45-57.
Flow cytometric determination of the number of bull sperm cells showed that the number of spermatozoa measured by light scattering may considerably differ from the actual number of sperm cells in the samples, depending on the proportion of contaminating particles, similar in size to sperm cells. No accurate information can be obtained from the sum of live and dead cells distinguished by means of double fluorescence staining, since a part of the sperm cell population is in a transitory state i.e. between the viable and dead states, so it cannot be stained by either dye. The number of spermatozoa in the sample can be determined very accurately if the sample is treated first with Nonidet-P-40 detergent then stained with propidium iodide. With this procedure the DNA content of each cell nucleus can be labeled and, through detecting the fluorescence signals, the actual sperm cell count of the sample can be determined with the accuracy of 95-98%.
流式细胞术测定公牛精子细胞数量表明，通过光散射测量的精子数量可能与样本中实际精子细胞数量有很大差异，这取决于与精子细胞大小相似的污染颗粒比例。通过双荧光染色区分的活细胞和死细胞总数无法获得准确信息，因为一部分精子细胞群体处于过渡状态，即在存活和死亡状态之间，所以两种染料都无法对其进行染色。如果样本先用诺乃洗涤剂 - P - 40处理，然后用碘化丙啶染色，样本中的精子数量可以非常准确地测定。通过这个程序，可以标记每个细胞核的DNA含量，并通过检测荧光信号，以95% - 98%的准确率确定样本中实际的精子细胞数量。
