Fluorescence biomembrane force probe: Concurrent quantitation of receptor-ligand kinetics and binding-induced intracellular signaling on a single cell

Yunfeng Chen, Baoyu Liu, Lining Ju, Jinsung Hong, Qinghua Ji, Wei Chen, Cheng Zhu

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

Abstract

Membrane receptor-ligand interactions mediate many cellular functions. Binding kinetics and downstream signaling triggered by these molecular interactions are likely affected by the mechanical environment in which binding and signaling take place. A recent study demonstrated that mechanical force can regulate antigen recognition by and triggering of the T-cell receptor (TCR). This was made possible by a new technology we developed and termed fluorescence biomembrane force probe (fBFP), which combines single-molecule force spectroscopy with fluorescence microscopy. Using an ultra-soft human red blood cell as the sensitive force sensor, a high-speed camera and real-time imaging tracking techniques, the fBFP is of ~1 pN (10-12 N), ~3 nm and ~0.5 msec in force, spatial and temporal resolution. With the fBFP, one can precisely measure single receptor-ligand binding kinetics under force regulation and simultaneously image binding-triggered intracellular calcium signaling on a single live cell. This new technology can be used to study other membrane receptor-ligand interaction and signaling in other cells under mechanical regulation.

Original languageEnglish (US)
Article numbere52975
Pages (from-to)1-13
Number of pages13
JournalJournal of Visualized Experiments
Volume2015
Issue number102
DOIs
StatePublished - Aug 4 2015
Externally publishedYes

Keywords

  • Adhesion
  • Bioengineering
  • Calcium
  • Fluorescence and force spectroscopy
  • Kinetics
  • Mechano-transduction
  • Receptor-ligand binding
  • Single cell
  • Single molecule

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

Fingerprint

Dive into the research topics of 'Fluorescence biomembrane force probe: Concurrent quantitation of receptor-ligand kinetics and binding-induced intracellular signaling on a single cell'. Together they form a unique fingerprint.

Cite this