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Bacterial cell wall nanoimaging by autoblinking microscopy.

Kevin Floc'h1, Françoise Lacroix1, Liliana Barbieri1

  • 1Univ. Grenoble Alpes, CEA, CNRS, IBS, F-38000, Grenoble, France.

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|September 21, 2018
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Summary
This summary is machine-generated.

Autofluorescence in bacteria, termed autoblinking, arises from fluorophores in growth media binding to the cell wall. This phenomenon enables super-resolution imaging of unlabeled bacteria and reveals cell wall dynamics.

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Area of Science:

  • Microscopy
  • Cell Biology
  • Biophysics

Background:

  • Spurious fluorescence, or autoblinking, is a common artifact in single-molecule localization microscopy (SMLM) of bacteria.
  • The precise origin and mechanism of autoblinking remain largely undetermined, hindering its potential use in imaging.

Purpose of the Study:

  • To elucidate the origin and mechanism of autoblinking in Deinococcus strains.
  • To investigate the potential of autoblinking as a super-resolution imaging technique for live, unlabeled bacteria.

Main Methods:

  • Systematic evaluation of environmental factors influencing autoblinking.
  • Analysis of photophysical properties of autoblinking molecules.
  • Application of autoblinking microscopy for imaging live Deinococcus radiodurans.
  • Combination with Photoactivated Localization Microscopy (PALM) for multicolor imaging.

Main Results:

  • Autoblinking originates from the transient binding of fluorophores from the growth medium to the bacterial cell wall, acting via a Point Accumulation for Imaging in Nanoscale Topography (PAINT) mechanism.
  • Autoblinking molecules preferentially associate with the plasma membrane.
  • Autoblinking microscopy successfully generated nanoscale images of live, unlabeled bacteria.
  • Super-resolution imaging revealed insights into bacterial septum formation and cell wall dynamics.

Conclusions:

  • Autoblinking is a PAINT mechanism driven by fluorophore-cell wall interactions, not an intrinsic bacterial property.
  • Autoblinking microscopy offers a novel, label-free super-resolution technique for studying bacterial cell biology.
  • This method can be combined with traditional SMLM techniques for advanced multicolor imaging applications.