Probing mechanotransduction in living cells by optical tweezers and FRET-based molecular force microscopy
LENS - European Laboratory for Non-Linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019, Sesto Fiorentino, Italy
2 Department of Physics and Astronomy, University of Florence, Via Sansone 1, 50019, Sesto Fiorentino, Italy
3 Department of Physics, University of Cyprus, P.O. Box 20537, 1678, Nicosia, Cyprus
Accepted: 24 February 2021
Published online: 12 March 2021
Cells sense mechanical signals and forces to probe the external environment and adapt to tissue morphogenesis, external mechanical stresses and a wide range of diverse mechanical cues. Here, we propose a combination of optical tools to manipulate single cells and measure the propagation of mechanical and biochemical signals inside them. Optical tweezers are used to trap microbeads that are used as handles to manipulate the cell plasma membrane; genetically encoded FRET-based force sensors inserted in F-actin and alpha-actinin are used to measure the propagation of mechanical signals to the cell cytoskeleton, while fluorescence microscopy with single-molecule sensitivity can be used with a huge array of biochemical and genetic sensors. We describe the details of the setup implementation, the calibration of the basic components and preliminary characterization of actin and alpha-actinin FRET-based force sensors.
© The Author(s) 2021
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