Fluctuation-Based Super-Resolution Traction Force Microscopy.
Fluctuation-based super-resolution microscopy
SACD
SRRF
live imaging
mechanobiology
traction force microscopy
Journal
Nano letters
ISSN: 1530-6992
Titre abrégé: Nano Lett
Pays: United States
ID NLM: 101088070
Informations de publication
Date de publication:
08 04 2020
08 04 2020
Historique:
pubmed:
7
3
2020
medline:
26
1
2021
entrez:
7
3
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32142297
doi: 10.1021/acs.nanolett.9b04083
pmc: PMC7146861
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2230-2245Subventions
Organisme : Medical Research Council
ID : MR/K015826/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/T027924/1
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 203276/Z/16/Z
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/S507532/1
Pays : United Kingdom
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