{"title":"Measurement of Intracellular Ions by Flow Cytometry","authors":"Carl H. June, Jonni S. Moore","doi":"10.1002/0471142735.im0505s64","DOIUrl":null,"url":null,"abstract":"<p>The recent development of a number of new fluorescent probes makes it possible to measure the concentrations of various intracellular free ions in single living cells. Among these ions are calcium, magnesium, sodium, potassium, and hydrogen (pH). This unit describes flow cytometric protocols using the dyes Indo-1 AM, Fluo-3, and Fura Red AM to measure intracellular calcium concentration. Support protocols detail the use of calcium buffers to calibrate a flow cytometric calcium assay, and methods to facilitate dye loading; an alternate protocol describes the use of a spectrofluorimeter to measure intracellular calcium for those investigators without access to a flow cytometer.</p>","PeriodicalId":10733,"journal":{"name":"Current Protocols in Immunology","volume":"64 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142735.im0505s64","citationCount":"8","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Immunology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/0471142735.im0505s64","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Immunology and Microbiology","Score":null,"Total":0}
引用次数: 8
Abstract
The recent development of a number of new fluorescent probes makes it possible to measure the concentrations of various intracellular free ions in single living cells. Among these ions are calcium, magnesium, sodium, potassium, and hydrogen (pH). This unit describes flow cytometric protocols using the dyes Indo-1 AM, Fluo-3, and Fura Red AM to measure intracellular calcium concentration. Support protocols detail the use of calcium buffers to calibrate a flow cytometric calcium assay, and methods to facilitate dye loading; an alternate protocol describes the use of a spectrofluorimeter to measure intracellular calcium for those investigators without access to a flow cytometer.
近年来，一些新型荧光探针的发展使得测量单个活细胞内各种细胞内自由离子的浓度成为可能。这些离子中有钙、镁、钠、钾和氢(pH)。本单元描述了使用染料Indo-1 AM, Fluo-3和Fura Red AM测量细胞内钙浓度的流式细胞术方案。支持协议详细说明了使用钙缓冲液来校准流式细胞术钙测定，以及促进染料加载的方法;另一种方案描述了使用荧光光谱仪测量细胞内钙的研究人员没有获得流式细胞仪。
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