Sorting out bacterial viability with optical tweezers

被引:156
作者
Ericsson, M
Hanstorp, D
Hagberg, P
Enger, J
Nyström, T
机构
[1] Univ Gothenburg, Dept Cell & Mol Biol Microbiol, CMB Microbiol, S-40530 Gothenburg, Sweden
[2] Chalmers Univ Technol, Dept Phys, S-41296 Gothenburg, Sweden
关键词
D O I
10.1128/JB.182.19.5551-5555.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We have developed a method, using laser, optical tweezers and direct microscopic analysis of reproductive potential and membrane integrity, to assess single-cell viability in a stationary-phase Escherichia coli population. It is demonstrated here that a reduction in cell integrity, determined by using the fluorescent nucleic acid stain propidium iodide, correlated well with a reduction in cell proliferating potential during the stationary-phase period studied. Moreover. the same cells that exhibited reduced integrity were found to be the ones that failed to divide upon nutrient addition. A small but significant number of the intact cells (496 of 7,466 [6.6%]) failed to replicate. In other words, we did not find evidence for the existence of a large population of intact but nonculturable cells during the stationary-phase period studied but it is clear that reproductive ability can be lost prior to the loss of membrane integrity, In addition, about 1% of the stationary-phase cells were able to divide only once upon nutrient addition, and in a few cases, only one of the two cells produced by division was able to divide a second time, indicating that localized cell deterioration, inherited by only one of the daughters, mag occur. The usefulness of the optical trapping methodology in elucidating the mechanisms involved in stationary-phase-induced bacterial death and population heterogeneity is discussed.
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页码:5551 / 5555
页数:5
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