Polarized light scattering for rapid observation of bacterial size changes.

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.

[1]  A. L. Koch,et al.  Th size and shape of bacteria by light scattering measurements. , 1968, Biochimica et biophysica acta.

[2]  C. Woldringh,et al.  Morphological analysis of the division cycle of two Escherichia coli substrains during slow growth , 1977, Journal of bacteriology.

[3]  Z. Kam,et al.  Absorption and Scattering of Light by Small Particles , 1998 .

[4]  P. Wyatt Light scattering and the absolute characterization of macromolecules , 1993 .

[5]  W. Donachie,et al.  Cell length, cell growth and cell division , 1976, Nature.

[6]  H. E. Kubitschek,et al.  Cell volume increase in Escherichia coli after shifts to richer media , 1990, Journal of bacteriology.

[7]  O. Pierucci Dimensions of Escherichia coli at various growth rates: model for envelope growth , 1978, Journal of bacteriology.

[8]  P. Wyatt Differential light scattering: a physical method for identifying living bacterial cells. , 1968, Applied optics.

[9]  B. V. Bronk,et al.  Reproducibility and sensitivity of polarized light scattering for identifying bacterial suspensions. , 1989, Applied optics.

[10]  H. Bremer,et al.  Establishment of exponential growth after a nutritional shift-up in Escherichia coli B/r: accumulation of deoxyribonucleic acid, ribonucleic acid, and protein , 1977, Journal of bacteriology.

[11]  W. Bickel,et al.  Polarized light scattering from biological systems: A technique for cell differentiation , 1981 .

[12]  V R Stull Size Distribution of Bacterial Cells , 1972, Journal of bacteriology.