Open AccessLive-cell super-resolved PAINT imaging of piconewton cellular traction forces
Author(s) -
Joshua M. Brockman,
Hanquan Su,
Aaron T. Blanchard,
Yuxin Duan,
Travis A. Meyer,
M. Edward Quach,
Roxanne Glazier,
Alisina Bazrafshan,
Rachel L. Bender,
Anna Kellner,
Hiroaki Ogasawara,
Rong Ma,
Florian Schueder,
Brian G. Petrich,
Ralf Jungmann,
Renhao Li,
Alexa L. Mattheyses,
Yonggang Ke,
Khalid Salaita
Publication year - 2020
Publication title -
nature methods
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 19.469
H-Index - 318
eISSN - 1548-7105
pISSN - 1548-7091
DOI - 10.1038/s41592-020-0929-2
Subject(s) - force spectroscopy , tension (geology) , biophysics , live cell imaging , nanotechnology , temporal resolution , nanoscopic scale , image resolution , chemistry , materials science , atomic force microscopy , cell , optics , physics , biology , composite material , biochemistry , ultimate tensile strength
Despite the vital role of mechanical forces in biology, it still remains a challenge to image cellular force with sub-100-nm resolution. Here, we present tension points accumulation for imaging in nanoscale topography (tPAINT), integrating molecular tension probes with the DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) technique to map piconewton mechanical events with ~25-nm resolution. To perform live-cell dynamic tension imaging, we engineered reversible probes with a cryptic docking site revealed only when the probe experiences forces exceeding a defined mechanical threshold (~7-21 pN). Additionally, we report a second type of irreversible tPAINT probe that exposes its cryptic docking site permanently and thus integrates force history over time, offering improved spatial resolution in exchange for temporal dynamics. We applied both types of tPAINT probes to map integrin receptor forces in live human platelets and mouse embryonic fibroblasts. Importantly, tPAINT revealed a link between platelet forces at the leading edge of cells and the dynamic actin-rich ring nucleated by the Arp2/3 complex.