Stubb, A;
Laine, RF;
Miihkinen, M;
Hamidi, H;
Guzman, C;
Henriques, R;
Jacquemet, G;
(2020)
Fluctuation-Based Super-Resolution Traction Force Microscopy.
Nano Letters
, 20
(4)
pp. 2230-2245.
10.1021/acs.nanolett.9b04083.
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Abstract
Cellular mechanics play a crucial role in tissue homeostasis and are often misregulated in disease. Traction force microscopy is one of the key methods that has enabled researchers to study fundamental aspects of mechanobiology; however, traction force microscopy is limited by poor resolution. Here, we propose a simplified protocol and imaging strategy that enhances the output of traction force microscopy by increasing i) achievable bead density and ii) the accuracy of bead tracking. Our approach relies on super-resolution microscopy, enabled by fluorescence fluctuation analysis. Our pipeline can be used on spinning-disk confocal or widefield microscopes and is compatible with available analysis software. In addition, we demonstrate that our workflow can be used to gain biologically relevant information and is suitable for fast long-term live measurement of traction forces even in light-sensitive cells. Finally, using fluctuation-based traction force microscopy, we observe that filopodia align to the force field generated by focal adhesions.
| Type: | Article |
|---|---|
| Title: | Fluctuation-Based Super-Resolution Traction Force Microscopy |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1021/acs.nanolett.9b04083 |
| Publisher version: | https://doi.org/10.1021/acs.nanolett.9b04083 |
| Language: | English |
| Additional information: | This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
| Keywords: | Science & Technology, Physical Sciences, Technology, Chemistry, Multidisciplinary, Chemistry, Physical, Nanoscience & Nanotechnology, Materials Science, Multidisciplinary, Physics, Applied, Physics, Condensed Matter, Chemistry, Science & Technology - Other Topics, Materials Science, Physics, Fluctuation-based super-resolution microscopy, SACD, SRRF, traction force microscopy, live imaging, mechanobiology, CELL-ADHESION, FILOPODIA, BLINKING, IMAGE |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10097849 |
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