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Mechanically resolved imaging of bacteria using expansion microscopy.

Youngbin Lim1, Anthony L Shiver1, Margarita Khariton1

  • 1Department of Bioengineering, Stanford University, Stanford, California, United States of America.

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Expansion microscopy (μExM) physically expands bacterial cells, revealing unique cell wall properties for species differentiation. This novel imaging contrast enhances microbial community analysis in diverse biological contexts.

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Area of Science:

  • Microbiology
  • Microscopy
  • Biophysics

Background:

  • Imaging microbial communities is challenging due to similar species morphologies.
  • Existing spectral labeling techniques have limitations in resolving diverse bacteria.

Purpose of the Study:

  • To develop a novel imaging method for resolving bacterial species and states.
  • To utilize physical expansion as a phenotypic contrast for microbial imaging.

Main Methods:

  • Developed expansion microscopy (μExM) for physical expansion of bacterial cells before imaging.
  • Leveraged species- and condition-dependent cell wall expansion patterns as imaging contrast.
  • Applied μExM to analyze microbial communities in vitro and in vivo.

Main Results:

  • Expansion patterns correlate with cell wall properties, providing species-specific contrast.
  • μExM successfully distinguished bacterial species in a human gut commensal community.
  • Detected antibiotic-induced cell-envelope damage and phenotypic heterogeneity in pathogenic bacteria during macrophage infection.

Conclusions:

  • Expansion microscopy (μExM) offers a powerful, orthogonal contrast for resolving microbial diversity and states.
  • This method advances the study of host-microbe interactions and bacterial physiology.