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Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence (TIRF) Microscopy
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Biochemistry
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JoVE Journal Biochemistry
Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence (TIRF) Microscopy

Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence (TIRF) Microscopy

DOI:
10.3791/64074-v

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08:44 min

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July 20, 2022

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1Department of Biochemistry & Molecular Biology, and Department of Neuroscience & Physiology,State University of New York (SUNY) Upstate Medical University

Chapters

  • 00:04Introduction
  • 00:40Coating Cleaned Coverslips with mPEG- and Biotin-PEG-Silane
  • 01:21Assembling Imaging Flow Chambers
  • 02:50Conditioning of Perfusion Chambers
  • 03:47Microscope Preparation
  • 04:40Preparation of Protein Reaction Mixes
  • 06:02Image Actin and Microtubule Dynamics
  • 06:43Results: Example Measurements of Microtubules and Actin Filament Dynamics
  • 08:14Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol is a guide for visualizing dynamic actin and microtubules using an in vitro total internal fluorescence (TIRF) microscopy assay.

Tags

Keywords: ActinMicrotubuleTIRF MicroscopyPEG-SilaneBiotin-PEG-SilaneCover SlipImaging ChamberEpoxy ResinPerfusion ChamberConditioning SolutionsTotal Internal Reflection Fluorescence

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