Fluorescent probes and flow cytometry to assess rat sperm integrity and mitochondrial function
Fluorescent probes and flow cytometry to assess rat sperm integrity and mitochondrial function
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DOI：
10.1016/s0890-6238(00)00113-1
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发表时间：
2001-01-01
影响因子：
3.3
通讯作者：
Berger, T
中科院分区：
文献类型：
--
作者：
Gravance, CG;Garner, DL;Berger, T
通过流式细胞术对细胞完整性和线粒体功能进行荧光评估可以提供一种快速和精确的方法来确定大量精子的功能状态。在本研究中，用SYBR-14和PI评估大鼠精子活力，并用JC-1差异标记精子线粒体。使用不同比例的新鲜和冷冻精子制备不同活力的精子样本。SYBR-14染色的精子与预期的精子活力相关性良好（r = 0.98）。用JC-1染色的活动精子在中段呈现橙子，表明线粒体膜电位高，而：具有低膜电位的不活动精子染色为绿色。精子染色为橙子的百分比与预期的精子活力高度相关（r = 0.99）。应用特异性荧光探针的流式细胞术是检测大鼠精子质膜完整性和线粒体功能变化的有效方法。(C)2001 Elsevier Science Inc. All rights reserved.
Fluorescent assessment of cellular integrity and mitochondrial function by flow cytometry can provide a rapid and precise means of determining the functional status of large numbers of spermatozoa. In the present study, rat sperm viability was assessed with SYBR-14 and PI and sperm mitochondria were differentially labeled with JC-1. Sperm samples of variable viability were prepared using varying proportions of fresh and frozen spermatozoa. SYBR-14 stained sperm correlated well with expected sperm viability (r = 0.98). Motile sperm stained with JC-1 appeared orange in the midpiece indicating a high mitochondrial membrane potential whereas: immotile sperm with a low membrane potential stained green. The percentage of spermatozoa staining orange was highly correlated (r = 0.99) with expected sperm viability. Flow cytometry using specific fluorescent probes is a useful technique fur detecting changes in rat sperm plasma membrane integrity and mitochondrial function in large numbers of spermatozoa. (C) 2001 Elsevier Science Inc. All rights reserved.