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Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy
JoVE 杂志
免疫与感染
This content is Free Access.
JoVE 杂志 免疫与感染
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

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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

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

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