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Fluorescens Biomembrane Force Probe: Samtidig Kvantifisering av Receptor-ligand Kinetics og Binding-indusert intracellulære signal på en enkelt celle
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JoVE Journal Bioengineering
Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell

Fluorescens Biomembrane Force Probe: Samtidig Kvantifisering av Receptor-ligand Kinetics og Binding-indusert intracellulære signal på en enkelt celle

DOI:
10.3791/52975-v

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14:09 min

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August 04, 2015

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, , , , , ,
1Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences,Georgia Institute of Technology, 2Coulter Department of Biomedical Engineering,Georgia Institute of Technology, 3Charles Perkins Centre,The University of Sydney, 4Institute of Biophysics, Laboratory of RNA Biology,Chinese Academy of Sciences, 5University of Chinese Academy of Sciences, 6School of Medicine and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases,Zhejiang University

Chapters

  • 00:05Title
  • 02:28Human RBCs Isolation, Biotinylation and Osmolarity Adjustment
  • 04:07Preparation for Micropipettes and a Cell Chamber
  • 06:24BFP Experiment
  • 10:25Fluorescence BFP (FBFP) Experiment
  • 11:51Results: Representative T-cell Receptor and Peptide MHC Binding Kinetics Data & Calcium Fluorescence Data from the FBFP Experiment
  • 13:14Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a technique for concurrently measuring force-regulated single receptor-ligand binding kinetics and real-time imaging of calcium signaling in a single T lymphocyte.

Tags

Keywords: Fluorescence Biomembrane Force ProbeFBFPMembrane Receptor-ligand KineticsBinding-induced Intracellular SignalingSingle-molecule Force SpectroscopyFluorescence MicroscopyMechanical Force RegulationT-cell ReceptorTCRBinding KineticsCalcium Signaling

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