Limits of propidium iodide as a cell viability indicator for environmental bacteria

被引:217
作者
Shi, Lei [1 ]
Guenther, Susanne [1 ]
Huebschmann, Thomas [1 ]
Wick, Lukas Y. [1 ]
Harms, Hauke [1 ]
Mueller, Susann [1 ]
机构
[1] UFZ Helmholtz Ctr Environm Res, Dept Environm Microbiol, Helmholtz Ctr Environm Res Leipzig Halle, D-04318 Leipzig, Germany
关键词
multiparametric flow cytometry; bacterial viability; population dynamics; live/dead staining; propidium iodide;
D O I
10.1002/cyto.a.20402
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Viability measurements of individual bacteria are applied in various scopes of research and industry using approaches where propidium iodide (PI) serves as dead cell indicator. The reliability of PI uptake as a cell viability indicator for dead (PI permeable) and viable (PI impermeable) bacteria was tested using two soil bacteria, the gram(-) Sphingomonas sp. LB126 and the gram(+) Mycobacterium frederiksbergense LB501T. Bacterial proliferation activities observed via DAPI and Hoechst 33342 staining were linked to the energy charge and the proportion of dead cells as obtained by diOC(6) (3)-staining and Pl-uptake, respectively. Calibration and verification experiments were performed using batch cultures grown on different substrates. PI uptake depended on the physiological state of the bacterial cells. Unexpectedly, up to 40% of both strains were stained by PI during early exponential growth on glucose when compared to 2-5% of cells in the early stationary phase of growth. The results question the utility of PI as a universal indicator for the viability of (environmental) bacteria. It rather appears that in addition to nonviable cells, PI also stains growing cells of Sphingomonas sp. and M. frederiksbergense during a short period of their life cycle. (C) 2007 International Society for Analytical Cytology
引用
收藏
页码:592 / 598
页数:7
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