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Oligomerization dynamik af celleoverflade receptorer i levende celler ved total intern refleksion Fluorescens mikroskopi kombineret med tal og lysstyrke analyse
JoVE Journal
Biology
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JoVE Journal Biology
Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis
DOI:

10:43 min

November 06, 2019

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Chapters

  • 00:04Title
  • 01:48Sample Preparation
  • 02:38TIRF Imaging: Alignment of the Laser Line and Optimization of TIRF Illumination
  • 04:05TIRF Imaging: Capture of the Time Series
  • 05:46Number & Brightness (N&B): Quality Check of the Time Series
  • 06:34Number & Brightness (N&B): Computation of the B-values in Selected Region-of-interest (ROI)
  • 08:19Results: N&B Analysis and Kinetics of FGFR1 Oligomerization
  • 09:42Conclusion

Summary

Automatic Translation

Vi beskriver en billedbehandlings metode til bestemmelse af den gennemsnitlige oligomeriske tilstand af mEGFP-mærkede receptor-oligomerer induceret af ligand binding i plasma membranen af levende celler. Protokollen er baseret på total intern refleksion fluorescens (TIRF) mikroskopi kombineret med tal og lysstyrke (N & B) analyse.

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