Kali R. Iyer, Nicole Robbins, Leah E. Cowen
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Abstract
A technique to assess the ability of distinct Candida 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 Candida 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 Candida species. © 2020 Wiley Periodicals LLC
Basic Protocol 1 : Growth and sample preparation of stained Candida
Basic Protocol 2 : Quantitative measurement of fluorescence by flow cytometry
Alternate Protocol : Qualitative determination of fluorescence using microscopy
念珠菌种外排的流式细胞术测定
一种评估不同念珠菌菌株对外排底物的能力以及比较外排抑制剂有效性的技术，对于分析抗真菌药物耐药机制和抗真菌药物的作用方式非常重要。我们描述了一种测量念珠菌物种挤出荧光染料尼罗河红作为外排活性输出能力的方法。这包括将细胞暴露在尼罗红中，并使用流式细胞术定量细胞荧光，使大量样品能够在有限的时间内处理。该方案提供了一种简单而有效的方法来定量耐药念珠菌物种的外排。©2020 Wiley期刊有限公司基本方案1:染色念珠菌的生长和样品制备基本方案2:用流式细胞术定量测量荧光交替方案:用显微镜定性测定荧光
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