{"title":"Inter-and Intra-Laboratory Standardization of TUNEL Assay for Assessment of Sperm DNA Fragmentation","authors":"Sajal Gupta, Rakesh Sharma, Ashok Agarwal","doi":"10.1002/cptx.37","DOIUrl":null,"url":null,"abstract":"<p>The functional aspects of sperm activity such as sperm chromatin integrity and ability to fertilize cannot be characterized by routine semen parameters. Men with unexplained infertility and idiopathic infertility, as well as men with normozoospermic semen profiles, show high DNA fragmentation. Molecular anomalies in the sperm can be detected by a sperm DNA fragmentation (SDF) assay which can be used in adjunct to conventional semen analysis. While the sperm chromatin structure assay (SCSA) remains the “gold standard,” the TUNEL assay using flow cytometry is becoming popular among the different tests that are currently available to measure sperm DNA fragmentation. In this unit, we describe the inter-laboratory and intra-laboratory standardization of the TUNEL assay using a benchtop cytometer. The article also provides a step-by-step protocol for measuring sperm DNA fragmentation using the TUNEL assay and a bench-top flow cytometer, and also points out the inherent challenges with this test. © 2017 by John Wiley & Sons, Inc.</p>","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"74 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.37","citationCount":"35","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in toxicology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cptx.37","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 35
Abstract
The functional aspects of sperm activity such as sperm chromatin integrity and ability to fertilize cannot be characterized by routine semen parameters. Men with unexplained infertility and idiopathic infertility, as well as men with normozoospermic semen profiles, show high DNA fragmentation. Molecular anomalies in the sperm can be detected by a sperm DNA fragmentation (SDF) assay which can be used in adjunct to conventional semen analysis. While the sperm chromatin structure assay (SCSA) remains the “gold standard,” the TUNEL assay using flow cytometry is becoming popular among the different tests that are currently available to measure sperm DNA fragmentation. In this unit, we describe the inter-laboratory and intra-laboratory standardization of the TUNEL assay using a benchtop cytometer. The article also provides a step-by-step protocol for measuring sperm DNA fragmentation using the TUNEL assay and a bench-top flow cytometer, and also points out the inherent challenges with this test. © 2017 by John Wiley & Sons, Inc.
用于评估精子DNA片段的TUNEL测定的实验室间和实验室内标准化
精子活动的功能方面,如精子染色质完整性和受精能力不能通过常规精液参数来表征。不明原因不孕症和特发性不孕症的男性,以及精子正常的男性,显示出高的DNA片段化。精子中的分子异常可以通过精子DNA碎片(SDF)测定来检测,该测定可用于辅助常规精液分析。虽然精子染色质结构分析(SCSA)仍然是“金标准”,但使用流式细胞术的TUNEL分析在目前可用于测量精子DNA片段的不同测试中越来越受欢迎。在本单元中,我们描述了使用台式细胞仪进行TUNEL测定的实验室间和实验室内标准化。本文还提供了使用TUNEL测定和台式流式细胞仪测量精子DNA片段的逐步方案,并指出了该测试的固有挑战。©2017 by John Wiley &儿子,Inc。
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