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{"title":"Detection and Quantification of Mitochondrial Fusion Using Imaging Flow Cytometry","authors":"Aldo Nascimento, Joanne Lannigan, David Kashatus","doi":"10.1002/cpcy.26","DOIUrl":null,"url":null,"abstract":"<p>Mitochondria are dynamic organelles that perform several vital cellular functions. Requisite for these functions are mitochondrial fusion and fission. Despite the increasing importance of mitochondrial dynamics in a range of cellular processes, there exist limited methods for robust quantification of mitochondrial fission and fusion. Currently, the most widely used method to measure mitochondrial fusion is the polyethylene glycol (PEG) fusion assay. While this assay can provide useful information regarding fusion activity, the reliance on manual selection of rare fusion events is time consuming and may introduce selection bias. By utilizing the image-capture features and colocalization analysis of imaging flow cytometry in combination with the PEG fusion assay, we are able to develop a high-throughput method to detect and quantify mitochondrial fusion activity. © 2017 by John Wiley & Sons, Inc.</p>","PeriodicalId":11020,"journal":{"name":"Current Protocols in Cytometry","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2017-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpcy.26","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cytometry","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpcy.26","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Health Professions","Score":null,"Total":0}
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Abstract
Mitochondria are dynamic organelles that perform several vital cellular functions. Requisite for these functions are mitochondrial fusion and fission. Despite the increasing importance of mitochondrial dynamics in a range of cellular processes, there exist limited methods for robust quantification of mitochondrial fission and fusion. Currently, the most widely used method to measure mitochondrial fusion is the polyethylene glycol (PEG) fusion assay. While this assay can provide useful information regarding fusion activity, the reliance on manual selection of rare fusion events is time consuming and may introduce selection bias. By utilizing the image-capture features and colocalization analysis of imaging flow cytometry in combination with the PEG fusion assay, we are able to develop a high-throughput method to detect and quantify mitochondrial fusion activity. © 2017 by John Wiley & Sons, Inc.
成像流式细胞术检测和定量线粒体融合
线粒体是动态的细胞器,执行一些重要的细胞功能。这些功能的必要条件是线粒体的融合和裂变。尽管线粒体动力学在一系列细胞过程中越来越重要,但线粒体裂变和融合的可靠量化方法有限。目前,最广泛使用的方法来测量线粒体融合是聚乙二醇(PEG)融合试验。虽然该分析可以提供有关融合活动的有用信息,但依赖于人工选择罕见的融合事件是耗时的,并且可能引入选择偏差。通过利用成像流式细胞术的图像捕获特性和共定位分析,结合PEG融合测定,我们能够开发出一种高通量方法来检测和量化线粒体融合活性。©2017 by John Wiley &儿子,Inc。
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