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Biophysical Measurements of Bacterial Cell Shape

  • Jeffrey P. Nguyen
  • Benjamin P. Bratton
  • Joshua W. Shaevitz
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1440)

Abstract

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.

Key words

Cell shape Fluorescent microscopy Active contours Bacteria 3D shape Deconvolution Point spread function 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jeffrey P. Nguyen
    • 1
    • 2
  • Benjamin P. Bratton
    • 1
    • 3
  • Joshua W. Shaevitz
    • 1
    • 2
  1. 1.Lewis-Sigler Institute for Integrative GenomicsPrinceton UniversityPrincetonUSA
  2. 2.Department of PhysicsPrinceton UniversityPrincetonUSA
  3. 3.Department of Molecular BiologyPrinceton UniversityPrincetonUSA

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