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A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

DOI:
10.3791/3892-v

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16:10 min

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March 22, 2012

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, ,
1Laboratory of Cellular and Molecular Immunology,National Institute of Allergy and Infectious Diseases, National Institutes of Health

Chapters

  • 00:05Title
  • 02:59Transfection Using Amaxa Mouse T Cell Nucleofector Kit
  • 06:07TIRF Setup
  • 10:24Imaging
  • 13:19Representative Results
  • 14:54Conclusion

Summary

Automatic Translation

Automatic Translation

The compartmentalization of proteins either within the plasma membrane or into intracellular locations is one regulatory mechanism that can greatly influence signaling outcomes; hence, to understand signaling it is important to study the spatial and temporal behavior of the proteins involved. We describe here a TIRF microscopy based system to study signal transduction in T cells, but is broadly applicable.

Tags

TIRF MicroscopyReal-time ImagingTCRSignaling ProteinsAntigen Presenting CellsPMHC ProteinsTCR ComplexCD3 DimersSrc Family KinaseLckSyk Family KinaseZap70Adapter ProteinsLATSLP76Proximal Signaling ComplexCalcium-dependent Transcription Factor ActivationGene Expression ProgramsT Cell DifferentiationAntigen PotencyCo-stimulatory ReceptorsCo-inhibitory ReceptorsChemokine ReceptorsCytokine Receptors

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