A1 Refereed original research article in a scientific journal
Fluctuation-Based Super-Resolution Traction Force Microscopy
Authors: Aki Stubb, Romain F. Laine, Mitro Miihkinen, Hellyeh Hamidi, Camilo Guzmán, Ricardo Henriques, Guillaume Jacquemet, Johanna Ivaska
Publisher: AMER CHEMICAL SOC
Publication year: 2020
Journal: Nano Letters
Journal name in source: NANO LETTERS
Journal acronym: NANO LETT
Volume: 20
Issue: 4
First page : 2230
Last page: 2245
Number of pages: 16
ISSN: 1530-6984
eISSN: 1530-6992
DOI: https://doi.org/10.1021/acs.nanolett.9b04083
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/47353668
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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