Biophysical Journal 73: 500-506 (1997)
© 1997 the Biophysical Society

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Polarized light scattering for rapid observation of bacterial size changes.

Biomedical Instrumentation Center, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799, USA. wvandeme@barnabas.indwes.edu

ABSTRACT

The effect of changing growth conditions on the diameter of rod-shaped bacteria was studied in vivo with the use of polarized light scattering. The value of a ratio of scattering matrix elements was measured as a function of scattering angle at various times after nutritional "upshift" for two strains of Escherichia coli cells. The peak locations of the scattering function were calibrated against the diameter for rod-shaped bacteria. The peaks moved toward smaller angles as a function of time after upshift, indicating that the diameter was increasing. Under special conditions, substantial peak shifts occurred within a few minutes of growth condition change, indicating a rapid onset of growth in diameter. The rate of increase of the diameters after upshift was obtained from the angular shift of peak location. This rate was approximately 14 nm/min for E. coli K12 and approximately 9 nm/min for E. coli B/r at 37 degrees C. The rate of diameter increase is smaller at lower temperatures. Experiments with Bacillus megaterium showed that any diameter change after nutritional upshift at 37 degrees C is limited to at most a very small increase, at least for the strain and medium tested.