Biophysical Measurements of Bacterial Cell Shape.
A bacteria's shape plays a large role in determining its mechanism of motility, energy requirements, and ability to avoid predation. Although it is a major factor in cell fitness, little is known about how cell shape is determined or maintained. These problems are made worse by a lack of accurate methods to measure cell shape in vivo, as current methods do not account for blurring artifacts introduced by the microscope. Here, we introduce a method using 2D active surfaces and forward convolution with a measured point spread function to measure the 3D shape of different strains of E. coli from fluorescent images. Using this technique, we are also able to measure the distribution of fluorescent molecules, such as polymers, on the cell surface. This quantification of the surface geometry and fluorescence distribution allow for a more precise measure of 3D cell shape and is a useful tool for measuring protein localization and the mechanisms of bacterial shape control.
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.|Department of Physics, Princeton University, Princeton, NJ, USA.
Jeffrey P
JP
Nguyen
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.|Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
Benjamin P
BP
Bratton
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.|Department of Physics, Princeton University, Princeton, NJ, USA.
Joshua W
JW
Shaevitz
D000465:Algorithms; D055592:Biophysical Phenomena; D004926:Escherichia coli / Q000502:physiology*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence
3D shape; Active contours; Bacteria; Cell shape; Deconvolution; Fluorescent microscopy; Point spread function