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Oligomerization dynamik af celleoverflade receptorer i levende celler ved total intern refleksion Fluorescens mikroskopi kombineret med tal og lysstyrke analyse
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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

Oligomerization dynamik af celleoverflade receptorer i levende celler ved total intern refleksion Fluorescens mikroskopi kombineret med tal og lysstyrke analyse

DOI:
10.3791/60398-v

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10:43 min

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November 06, 2019

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, , , ,
1Unit of Microscopy and Dynamic Imaging,Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain, 2Centro di Imaging Sperimentale,Ospedale San Raffaele, Milan, Italy, 3Unit of Gynecological Oncology Research,European Institute of Oncology IRCCS, Milan, Italy

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

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.

Tags

OligomerizationCell Surface ReceptorsTIRF MicroscopyNumber And Brightness AnalysisReceptor ClusteringFGFR1Fluorescence FluctuationLive Cell ImagingProtein OligomerizationCell Signaling

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