期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 102, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/52975
关键词
Bioengineering; Issue 102; single cell; single molecule; receptor-ligand binding; kinetics; fluorescence and force spectroscopy; adhesion; mechano-transduction; calcium
资金
- NIH [AI044902, AI077343, AI038282, HL093723, HL091020, GM096187, TW008753]
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 similar to 1 pN (10(-12) N), similar to 3 nm and similar to 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.
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