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{"title":"Cell Volume Measurements by Optical Transmission Microscopy","authors":"Michael A. Model","doi":"10.1002/0471142956.cy1239s72","DOIUrl":null,"url":null,"abstract":"<p>Cell volume is an important parameter in cell adaptation to anisosmotic stress, in the development of apoptosis and necrosis, and in the pathogenesis of several diseases. This unit describes a method for measuring the volume of adherent cells using a standard light microscope. A coverslip with attached cells is placed in a shallow chamber in a medium containing a strongly absorbing and cell-impermeant dye, Acid Blue 9. When such a sample is imaged in transmitted light at a wavelength of maximum dye absorption (630 nm), the resulting contrast quantitatively reflects cell thickness. Once the thickness is known at every point, the volume can be computed as well. Technical details, interpretation of data, and possible artifacts are discussed. Measurements in absolute units require knowledge of the absorption coefficient, and a similar procedure for the measurement of absorption coefficient is described. © 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.cy1239s72","citationCount":"12","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cytometry","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/0471142956.cy1239s72","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: 12
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Abstract
Cell volume is an important parameter in cell adaptation to anisosmotic stress, in the development of apoptosis and necrosis, and in the pathogenesis of several diseases. This unit describes a method for measuring the volume of adherent cells using a standard light microscope. A coverslip with attached cells is placed in a shallow chamber in a medium containing a strongly absorbing and cell-impermeant dye, Acid Blue 9. When such a sample is imaged in transmitted light at a wavelength of maximum dye absorption (630 nm), the resulting contrast quantitatively reflects cell thickness. Once the thickness is known at every point, the volume can be computed as well. Technical details, interpretation of data, and possible artifacts are discussed. Measurements in absolute units require knowledge of the absorption coefficient, and a similar procedure for the measurement of absorption coefficient is described. © 2015 by John Wiley & Sons, Inc.
用光学透射显微镜测量细胞体积
细胞体积是细胞适应异渗胁迫、细胞凋亡和坏死的发展以及多种疾病发病机制的重要参数。本单元描述了一种使用标准光学显微镜测量贴壁细胞体积的方法。将带有附着细胞的盖盖放在一个浅室中,介质中含有一种强吸收且不影响细胞的染料,酸性蓝9。当这样的样品在最大染料吸收波长(630 nm)的透射光下成像时,所得到的对比度定量地反映了细胞厚度。一旦知道了每个点的厚度,就可以计算出体积。讨论了技术细节、数据解释和可能的工件。以绝对单位测量需要了解吸收系数,并描述了测量吸收系数的类似程序。©2015 by John Wiley &儿子,Inc。
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