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{"title":"念珠菌种外排的流式细胞术测定","authors":"Kali R. Iyer, Nicole Robbins, Leah E. Cowen","doi":"10.1002/cpmc.121","DOIUrl":null,"url":null,"abstract":"<p>A technique to assess the ability of distinct <i>Candida</i> strains to efflux substrates, as well as to compare the effectiveness of efflux inhibitors, is important for analysis of antifungal drug resistance mechanisms and the mode of action of antifungals. We describe a method that measures the ability of <i>Candida</i> species to extrude the fluorescent dye Nile red as an output for efflux activity. This involves exposing cells to Nile red and using flow cytometry to quantify cellular fluorescence, enabling numerous samples to be processed in a limited time frame. This protocol provides a simple, yet effective method for quantifying efflux in drug-resistant <i>Candida</i> species. © 2020 Wiley Periodicals LLC</p><p><b>Basic Protocol 1</b>: Growth and sample preparation of stained <i>Candida</i></p><p><b>Basic Protocol 2</b>: Quantitative measurement of fluorescence by flow cytometry</p><p><b>Alternate Protocol</b>: Qualitative determination of fluorescence using microscopy</p>","PeriodicalId":39967,"journal":{"name":"Current Protocols in Microbiology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2020-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmc.121","citationCount":"2","resultStr":"{\"title\":\"Flow Cytometric Measurement of Efflux in Candida Species\",\"authors\":\"Kali R. Iyer, Nicole Robbins, Leah E. Cowen\",\"doi\":\"10.1002/cpmc.121\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>A technique to assess the ability of distinct <i>Candida</i> strains to efflux substrates, as well as to compare the effectiveness of efflux inhibitors, is important for analysis of antifungal drug resistance mechanisms and the mode of action of antifungals. We describe a method that measures the ability of <i>Candida</i> species to extrude the fluorescent dye Nile red as an output for efflux activity. This involves exposing cells to Nile red and using flow cytometry to quantify cellular fluorescence, enabling numerous samples to be processed in a limited time frame. This protocol provides a simple, yet effective method for quantifying efflux in drug-resistant <i>Candida</i> species. © 2020 Wiley Periodicals LLC</p><p><b>Basic Protocol 1</b>: Growth and sample preparation of stained <i>Candida</i></p><p><b>Basic Protocol 2</b>: Quantitative measurement of fluorescence by flow cytometry</p><p><b>Alternate Protocol</b>: Qualitative determination of fluorescence using microscopy</p>\",\"PeriodicalId\":39967,\"journal\":{\"name\":\"Current Protocols in Microbiology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-10-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/cpmc.121\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Protocols in Microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.121\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.121","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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