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{"title":"Flow Cytometry of Murine Spermatocytes","authors":"Valeriya Gaysinskaya, Alex Bortvin","doi":"10.1002/0471142956.cy0744s72","DOIUrl":null,"url":null,"abstract":"<p>Protocols for purification of murine male germ cells by FACS based on Hoechst 33342 (Ho342) dye staining have been reported and optimized. However, the protocols are often challenging to follow, partly due to difficulties related to sample preparation, instrument parameters, data display, and selection strategies. In addition, troubleshooting of flow cytometry experiments usually requires some fluency in technical principles and instrument specifications and settings. This unit describes setup and procedures for analysis and sorting of male meiotic prophase I (MPI) cells and other germ cells. Included are procedures that guide data acquisition, display, gating, and back-gating critical for optimal data visualization and cell sorting. Additionally, a flow cytometry analysis of spermatogenesis-defective testis is provided to illustrate the applicability of the technique to the characterization and purification of cells from mutant testis. © 2015 by John Wiley & Sons, Inc.</p>","PeriodicalId":11020,"journal":{"name":"Current Protocols in Cytometry","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2015-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142956.cy0744s72","citationCount":"24","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cytometry","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/0471142956.cy0744s72","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Health Professions","Score":null,"Total":0}
引用次数: 24
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Abstract
Protocols for purification of murine male germ cells by FACS based on Hoechst 33342 (Ho342) dye staining have been reported and optimized. However, the protocols are often challenging to follow, partly due to difficulties related to sample preparation, instrument parameters, data display, and selection strategies. In addition, troubleshooting of flow cytometry experiments usually requires some fluency in technical principles and instrument specifications and settings. This unit describes setup and procedures for analysis and sorting of male meiotic prophase I (MPI) cells and other germ cells. Included are procedures that guide data acquisition, display, gating, and back-gating critical for optimal data visualization and cell sorting. Additionally, a flow cytometry analysis of spermatogenesis-defective testis is provided to illustrate the applicability of the technique to the characterization and purification of cells from mutant testis. © 2015 by John Wiley & Sons, Inc.
小鼠精母细胞流式细胞术
本文报道并优化了基于Hoechst 33342 (Ho342)染色的FACS纯化小鼠雄性生殖细胞的方法。然而，这些方案往往具有挑战性，部分原因是与样品制备、仪器参数、数据显示和选择策略相关的困难。此外，流式细胞术实验的故障排除通常需要对技术原理和仪器规格和设置有一定的了解。本单元描述了雄性减数分裂前期I (MPI)细胞和其他生殖细胞的分析和分选的设置和程序。包括指导数据采集、显示、门控和反向门控的过程，这些过程对于优化数据可视化和单元格排序至关重要。此外，对精子发生缺陷的睾丸进行了流式细胞术分析，以说明该技术对突变睾丸细胞的表征和纯化的适用性。©2015 by John Wiley &儿子,Inc。
本文章由计算机程序翻译，如有差异，请以英文原文为准。
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