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Single-Molecule Tracking Microscopy – A Tool for Determining the Diffusive States of Cytosolic Molecules
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Biochemistry
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JoVE Journal Biochemistry
Single-Molecule Tracking Microscopy – A Tool for Determining the Diffusive States of Cytosolic Molecules

Single-Molecule Tracking Microscopy – A Tool for Determining the Diffusive States of Cytosolic Molecules

DOI:
10.3791/59387-v

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00:10 min

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September 05, 2019

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,
1Department of Chemistry,University of Virginia, 2Department of Molecular Physiology & Biological Physics,University of Virginia School of Medicine

Chapters

  • 00:04Title
  • 00:35Microscope Preparation
  • 02:33Y. enterocolitica Culture Preparation
  • 03:33Data Acquisition
  • 05:54Data Processing
  • 06:45Data Post-processing – Overlay of Outlines
  • 07:32Results: Determining the Diffusive States of YscQ
  • 09:25Conclusion

Summary

Automatic Translation

Automatic Translation

3D single-molecule localization microscopy is utilized to probe the spatial positions and motion trajectories of fluorescently labeled proteins in living bacterial cells. The experimental and data analysis protocol described herein determines the prevalent diffusive behaviors of cytosolic proteins based on pooled single-molecule trajectories.

Tags

Keywords: Single-molecule Tracking3D TrackingDiffusive StatesCytosolic MoleculesFluorescence MicroscopyAgarose PadYersinia EnterocoliticaCell VolumeZ-calibrationMATLABHCImage Live

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