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流式细胞术用于纯培养物中细菌浓度快速计数的验证

Validation of flow cytometry for rapid enumeration of bacterial concentrations in pure cultures.

作者信息

Pinder A C, Purdy P W, Poulter S A, Clark D C

机构信息

Department of Chemical Physics, AFRC Institute of Food Research, Norwich Laboratory, UK.

出版信息

J Appl Bacteriol. 1990 Jul;69(1):92-100. doi: 10.1111/j.1365-2672.1990.tb02916.x.

Abstract

Flow cytometry was investigated as a rapid detection and counting method for bacteria in pure cultures. A simple two-parameter detection scheme was employed: particle size was measured by forward angle light scatter and nucleic acid content by fluorescence of the DNA/RNA-binding dye ethidium bromide. The technique gave results that correlated exceptionally well with conventional plate counting for four species of bacteria, and concentrations in the range 10(2) to 10(7) cfu/ml. Cytometric counts were obtained in a few minutes, as compared with 2 d required for the plate counts. Under ideal conditions, each bacterial species examined exhibited a characteristic 'signature' on the cytometer, which could be explained by its known properties and morphology.

摘要

流式细胞术作为一种用于纯培养物中细菌的快速检测和计数方法进行了研究。采用了一种简单的双参数检测方案:通过前向角光散射测量颗粒大小,通过DNA/RNA结合染料溴化乙锭的荧光测量核酸含量。该技术所得结果与四种细菌的传统平板计数法以及浓度范围在10²至10⁷ cfu/ml时的结果相关性极佳。流式细胞术计数在几分钟内即可完成,而平板计数则需要2天时间。在理想条件下,所检测的每种细菌在细胞仪上都呈现出特征性的“信号”,这可以通过其已知特性和形态来解释。

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