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Probing mechanotransduction in living cells by optical tweezers and FRET-based molecular force microscopy

M. Sergides, L. Perego, T. Galgani, C. Arbore, F.S. Pavone, M Capitanio
doi: https://doi.org/10.1101/2021.01.24.427943
M. Sergides
1LENS - European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy
2Department of Physics and Astronomy, University of Florence, Via Sansone 1, 50019 Sesto Fiorentino, Italy
3Department of Physics, University of Cyprus, P.O. Box 20537, Nicosia, 1678, Cyprus
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L. Perego
1LENS - European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy
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T. Galgani
2Department of Physics and Astronomy, University of Florence, Via Sansone 1, 50019 Sesto Fiorentino, Italy
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C. Arbore
1LENS - European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy
2Department of Physics and Astronomy, University of Florence, Via Sansone 1, 50019 Sesto Fiorentino, Italy
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F.S. Pavone
1LENS - European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy
2Department of Physics and Astronomy, University of Florence, Via Sansone 1, 50019 Sesto Fiorentino, Italy
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M Capitanio
1LENS - European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy
2Department of Physics and Astronomy, University of Florence, Via Sansone 1, 50019 Sesto Fiorentino, Italy
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  • For correspondence: capitanio@lens.unifi.it
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Abstract

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.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 24, 2021.
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Probing mechanotransduction in living cells by optical tweezers and FRET-based molecular force microscopy
M. Sergides, L. Perego, T. Galgani, C. Arbore, F.S. Pavone, M Capitanio
bioRxiv 2021.01.24.427943; doi: https://doi.org/10.1101/2021.01.24.427943
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Probing mechanotransduction in living cells by optical tweezers and FRET-based molecular force microscopy
M. Sergides, L. Perego, T. Galgani, C. Arbore, F.S. Pavone, M Capitanio
bioRxiv 2021.01.24.427943; doi: https://doi.org/10.1101/2021.01.24.427943

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