龙胆紫染色用于分析精子存活率的方法学研究

A methyl violet staining method for sperm analysis on survival rate

目的:探讨龙胆紫染色区分存活精子和死亡精子可能性,以提供一种判定精子存活率的新方法,为男性精液检测和质量评价提供依据。方法:采用不同浓度的龙胆紫溶液对精液样本染色,在光学显微镜下观察染色结果,讨论区分存活精子和死亡精子的标准,并计数一定数量的总精子数,计算存活率,并将计算结果与标准方法比较。结果:经过龙胆紫溶液(0.05%,M/V)染色后在显微镜下存活精子呈现淡蓝色,而死亡精子呈现深紫色,计算获得的存活率与伊红染色法(0.5%,M/V)的结果比较差异统计学意义(t=0.862,双侧P=0.403)。结论:一定浓度的龙胆紫染色后能够清楚地区分存活精子和死亡精子,该法获得的精子存活率结果可靠,龙胆紫染色法可以作为一种新的分析精子存活率的方法。

Objective： To discuss whether methyl violet staining could distinguish between live and dead sperm, and in turn, to provide a new method for determining sperm survival and service male semen testing and the quality evalua- tion. Methods： Semen samples were stained with different concentrations of methyl violet solution and the staining re- sults were observed under optical microscope to distinguish the live sperm and the dead sperm. The amount of total sperm was counted and the survival rate was calculated. The same semen samples were also analyzed with eosin stai ning, and the survival rates from two methods were compared statistically. Results： After methyl violet solution stai- ning survived sperm was mainly light blue under the microscope, and dead sperm was mainly purple, the survival rate calculated was of no significant difference comparing with that of eosin staining method. Conclusion： The results of this study prove that a certain concentration of methyl violet staining can clearly distinguish the live sperm and the dead one, further the sperm survival rate of methyl violet could be reliable, and can be used as a new analysis of sperm sur- vival evaluation.