A SUPRAVITAL STAINING TECHNIQUE FOR HONEY-BEE SPERMATOZOA

被引：31

作者：

LOCKE, SJ

PENG, YS

CROSS, NL

机构：

[1] UNIV CALIF DAVIS,DEPT ENTOMOL,DAVIS,CA 95616

[2] UNIV CALIF DAVIS,DEPT OBSTET & GYNECOL,DAVIS,CA 95616

来源：

PHYSIOLOGICAL ENTOMOLOGY | 1990年 / 15卷 / 02期

关键词：

Apis mellifera; Hoechst; 33342; honey bee; insect; propidium iodide; semen quality; Spermatozoa; supravital stains;

D O I：

10.1111/j.1365-3032.1990.tb00506.x

中图分类号：

Q96 [昆虫学];

学科分类号：

摘要：

ABSTRACT A new supravital staining technique is described for honey bee, Apis mellifera L., spermatozoa using the fluorochromes, propidium iodide and Hoechst 33342 (H342), a bis‐benzimidazole derivative. Propidium iodide binds to the DNA of sperm which lack membrane integrity and H342 binds to the DNA of all sperm. This assay is a simple and rapid method for determining the percentage nonviabiiity of a male honey bee's sperm. The recommended staining procedure is to incubate sperm in a solution of 5 μ.g/ml H342 and 10 μ.g/ml propidium iodide in modified Kiev solution for 15–20 min. After incubation, wet mounts of the sperm‐stain suspension are examined using fluorescence microscopy. Percentage nonviabiiity is determined by the ratio of propidium iodide stained sperm to H342 stained sperm. Copyright © 1990, Wiley Blackwell. All rights reserved

引用

页码：187 / 192

页数：6

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