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Immunology and Infection
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Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

DOI:
10.3791/56745-v

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08:55 min

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December 29, 2017

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, , ,
1Department of Tumor Immunology,Radboud University Medical Center

Chapters

  • 00:05Title
  • 00:37Preparation of the Microscopy Samples
  • 01:49Recording of the FLIM Data
  • 03:34Conversion of the Photon Recordings to FLIM Images
  • 04:19Fitting of Fluorescence Lifetime Histograms for Whole-cell FLIM Analysis
  • 06:03Results: Syntaxin 4 and VAMP3 Interaction in Live Cell Visualized by FRET-FLIM
  • 07:43Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a new method allowing for the quantitative visualization of complex formation of SNARE proteins, based on Förster resonance energy transfer, and fluorescence lifetime imaging microscopy.

Tags

Keywords: SNAREFLIMMembrane TraffickingHeLa CellsSyntaxin-4VAMP3Syntaxin-3MCitrineMCherryFluorescence Lifetime Imaging MicroscopyConfocal MicroscopyInstrument Response Function

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