A supravital staining technique for honey bee spermatozoa |
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Authors: | SARAH J LOCKE YING-SHIN PENG NICHOLAS L CROSS |
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Institution: | Department of Entomology;Department of Entomology, University of California, Davis, CA 95616, U.S.A |
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Abstract: | 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. |
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Keywords: | Spermatozoa insect honey bee semen quality supravital stains Hoechst 33342 propidium iodide Apis mellifera |
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