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Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies
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Detecting Bacterial Surface Organelles on Single Cells Using Optical Tweezers.

Johan Zakrisson1, Bhupender Singh1, Pontus Svenmarker1

  • 1Department of Physics, and ‡Department of Chemistry, Umeå University , 901 87 Umeå, Sweden.

Langmuir : the ACS Journal of Surfaces and Colloids
|April 19, 2016
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method to detect adhesion pili on single bacteria. This technique uses optical tweezers to measure fluid drag, distinguishing piliated from non-piliated cells in real-time for further study.

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Area of Science:

  • Microbiology
  • Biophysics
  • Cell Biology

Background:

  • Bacterial surface organelles, like adhesion pili, are crucial for intercellular communication, motility, and adhesion.
  • Gram-negative bacteria use adhesion pili for attachment to host cells, but their small size prevents detection by standard light microscopy.
  • Current methods lack a fast, simple way to identify pili on live single bacterial cells.

Purpose of the Study:

  • To develop a novel, rapid, and non-destructive method for detecting adhesion pili on individual bacterial cells.
  • To enable real-time differentiation between piliated and non-piliated bacteria for subsequent live-cell studies.

Main Methods:

  • Imaging bacteria to measure cell size and predict fluid drag using an analytical model.
  • Trapping single bacteria with optical tweezers and oscillating the sample to measure effective fluid drag.
  • Comparing predicted fluid drag with measured fluid drag to infer the presence of pili.

Main Results:

  • The method successfully detected the presence of pili by comparing predicted and measured fluid drag.
  • Verified the protocol using polymer-coated silica microspheres and Escherichia coli expressing adhesion pili.
  • Demonstrated real-time, within-seconds capability to distinguish piliated from non-piliated bacteria.

Conclusions:

  • This optical tweezer-based fluid drag measurement offers a fast and simple approach to identify pili on single bacterial cells.
  • The developed protocol supports live-cell studies by enabling rapid identification of piliated bacteria.
  • This technique advances the study of bacterial adhesion, biofilm formation, and infection dynamics.