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Journal of Bacteriology, October 2000, p. 5551-5555, Vol. 182, No. 19
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Sorting Out Bacterial Viability with Optical Tweezers

M. Ericsson,1 D. Hanstorp,1 P. Hagberg,1 J. Enger,1 and T. Nyström2,*

Department of Physics, Chalmers University of Technology and Göteborg University,1 and Department of Cell and Molecular Biology-Microbiology, Göteborg University,2 Göteborg, Sweden

Received 15 March 2000/Accepted 6 July 2000

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, may occur. The usefulness of the optical trapping methodology in elucidating the mechanisms involved in stationary-phase-induced bacterial death and population heterogeneity is discussed.

* Corresponding author. Mailing address: CMB-Microbiology, Göteborg University, Box 462, 405 30 Göteborg, Sweden. Phone: (46) 31 773 2582. Fax: (46) 31 773 2599. E-mail: thomas.nystrom{at}gmm.gu.se.

Journal of Bacteriology, October 2000, p. 5551-5555, Vol. 182, No. 19
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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