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{"title":"Measurement of Intracellular Ions by Flow Cytometry","authors":"Avery D. Posey, Jr., Omkar U. Kawalekar, Carl H. June","doi":"10.1002/0471142956.cy0908s72","DOIUrl":null,"url":null,"abstract":"<p>Using flow cytometry, single-cell measurements of calcium can be made on isolated populations identified by one or more phenotypic characteristics. Most earlier techniques for measuring cellular activation parameters determined the mean value for a population of cells, which did not permit optimal resolution of the responses. The flow cytometer is particularly useful for this purpose because it can measure ion concentrations in large numbers of single cells and thereby allows ion concentration to be correlated with other parameters such as immunophenotype and cell cycle stage. A limitation of flow cytometry, however, is that it does not permit resolution of certain complex kinetic responses such as cellular oscillatory responses. This unit describes the preparation of cells, including labeling with antibodies and with calcium probes, and discusses the principles of data analysis and interpretation. © 2015 by John Wiley & Sons, Inc.</p>","PeriodicalId":11020,"journal":{"name":"Current Protocols in Cytometry","volume":"72 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2015-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142956.cy0908s72","citationCount":"29","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cytometry","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/0471142956.cy0908s72","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: 29
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Abstract
Using flow cytometry, single-cell measurements of calcium can be made on isolated populations identified by one or more phenotypic characteristics. Most earlier techniques for measuring cellular activation parameters determined the mean value for a population of cells, which did not permit optimal resolution of the responses. The flow cytometer is particularly useful for this purpose because it can measure ion concentrations in large numbers of single cells and thereby allows ion concentration to be correlated with other parameters such as immunophenotype and cell cycle stage. A limitation of flow cytometry, however, is that it does not permit resolution of certain complex kinetic responses such as cellular oscillatory responses. This unit describes the preparation of cells, including labeling with antibodies and with calcium probes, and discusses the principles of data analysis and interpretation. © 2015 by John Wiley & Sons, Inc.
流式细胞术测定细胞内离子
使用流式细胞术,钙的单细胞测量可以在一个或多个表型特征鉴定的分离群体。大多数早期的测量细胞激活参数的技术确定了细胞群的平均值,这并不能保证反应的最佳分辨率。流式细胞仪特别适用于此目的,因为它可以测量大量单细胞中的离子浓度,从而使离子浓度与其他参数(如免疫表型和细胞周期阶段)相关。然而,流式细胞术的一个局限性是,它不允许解决某些复杂的动力学反应,如细胞振荡反应。本单元描述了细胞的制备,包括用抗体和钙探针标记,并讨论了数据分析和解释的原理。©2015 by John Wiley &儿子,Inc。
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