Fluctuation-Based Super-Resolution Traction Force Microscopy.

Nano Letters
Aki StubbJohanna Ivaska

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.

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Citations

Dec 17, 2020·Sensors·Ying Tu, Xuefeng Wang
Dec 3, 2020·Frontiers in Bioengineering and Biotechnology·Nicolás Andrés SaffiotiDiego Pallarola
Mar 16, 2021·Frontiers in Cell and Developmental Biology·Marco Fritzsche
Apr 17, 2021·Nature Communications·Lucas von ChamierRicardo Henriques
Apr 14, 2021·Nature Communications·Liliana BarbieriMarco Fritzsche
Apr 14, 2021·Nature Communications·Di LiMarco Fritzsche
Apr 23, 2021·ACS Applied Materials & Interfaces·Ajinkya GhagreAllen Ehrlicher
Apr 27, 2021·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Kseniya KorobchevskayaMarco Fritzsche
May 6, 2021·Nature Communications·Thomas SchlichthaerleRalf Jungmann
Jun 4, 2021·Nano Letters·Nafsika ChalaAldo Ferrari
May 19, 2021·Cellular Signalling·Yasaswi Gayatri Mishra, Bramanandam Manavathi
Aug 21, 2021·Micron : the International Research and Review Journal for Microscopy·Małgorzata LekkaJoanna Zemła
Aug 29, 2021·Polymers·Zhuonan Yu, Kuo-Kang Liu
Sep 8, 2021·Current Protocols·Nikhil Mittal, Sangyoon J Han
Oct 27, 2021·Journal of the American Chemical Society·Ming-Ming ChenJing-Juan Xu

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Methods Mentioned

BETA
super-resolution microscopy
fluorescence imaging
force measurements
fluorescence-activated cell sorter

Software Mentioned

Noise2VOID
NanoJ SQUIRREL
PlotsOfData
Fiji
SQUIRREL
NanoJ
LiveSRRF
Core
ImageJ
Jupyter Notebooks

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