荧光染色法检测虹鳟精子质量

Evaluation of the Sperm Quality of Rainbow Trout (Oncorhynchus mykiss) Using the Fluorescent Dye

本实验运用三种荧光染色方法对虹鳟精子的质量进行检测,并探讨荧光染色对虹鳟精子质量评价的可性及意义。在生殖季节采集虹鳟精液,分别进行Hoechst33342/碘化丙啶双染,吖啶橙染色及罗丹明123/碘化丙啶双染,然后将其制成涂片荧光显微镜下观察。经荧光染色的虹鳟精子质膜完整,DNA完整,可明显分辨出活体和死亡精子,线粒体膜功能正常精子的比率分别为90.7%、98.7%和96.4%。综上所述,结合三种检测方法可以简便快速地测定虹鳟精子的质量。

Three kinds of fluorescent dye were carried out to detect the quality of the spermatozoa of rainbow trout （Oncorhynchus mykiss）. The milt of rainbow trout were divided into three groups and stained with Hoechst33342/PI, acridine orange and Rhodamine 123/PI separately. Then they were observed under fluorescence microscope. The result showed that the rate of spermatozoa with unabroken plasmalemma was 90.7% , while that with double strand DNA and mitochondria membrane was 98.7% and 96.4% , respectively. Taken together, the quality of the rainbow trout spermatozoa could be determined by fluorescence staining conveniently and easily.