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Characterizing Single-Molecule Conformational Changes Under Shear Flow with Fluorescence Microscopy
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Bioengineering
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JoVE Journal Bioengineering
Characterizing Single-Molecule Conformational Changes Under Shear Flow with Fluorescence Microscopy

Characterizing Single-Molecule Conformational Changes Under Shear Flow with Fluorescence Microscopy

DOI:
10.3791/60784-v

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08:47 min

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January 25, 2020

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, , , , ,
1Department of Bioengineering,Lehigh University, 2Department of Materials Science and Engineering,Lehigh University, 3Department of Chemistry,Lehigh University, 4Department of Mechanical Engineering and Mechanics,Lehigh University

Chapters

  • 00:05Introduction
  • 01:18Preparation of PDMS Microfluidic Device
  • 03:28Microfluidic Device Surface Treatment
  • 05:05Fluorescence Microscopy
  • 07:18Results: Extension and Relaxation of VWF and Lambda DNA Molecules due to Shear Flow
  • 08:00Conclusion

Summary

Automatic Translation

Automatic Translation

We present a protocol for immobilizing single macromolecules in microfluidic devices and quantifying changes in their conformations under shear flow. This protocol is useful for characterizing the biomechanical and functional properties of biomolecules such as proteins and DNA in a flow environment.

Tags

Keywords: Single-moleculeConformational ChangesShear FlowFluorescence MicroscopyProteinDNABiophysicsMicrofluidic DevicePDMSSurface TreatmentReal-time VisualizationRheologyComplex Fluids

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