Turn-key mapping of cell receptor force orientation and magnitude using a commercial structured illumination microscope

• Type of Material: Article
• Authors: Aaron Blanchard, Georgia Institute of Technology; Dale J Combs, Emory University; Brockman M Joshua, Emory University; Joshua M Brockman, Georgia Institute of Technology; Anna V Kellner, Georgia Institute of Technology; Roxanne Glazier, Georgia Institute of Technology; Hanquan Su, Emory University; Rachel L Bender, Emory University; Alisina S Bazrafshan, Emory University; Wenchun Chen, Emory University; Renhao Li, Emory University; Alexa L Mattheyses, University of Alabama Birmingham; Khalid Salaita, Emory University
• Language: English
• Date: 2021-08-03
• Publisher: NATURE PORTFOLIO
• Publication Version: Final Published Version
• Copyright Statement: © The Author(s) 2021, corrected publication 2021
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1038/s41467-021-24602-x
• Title of Journal or Parent Work: NATURE COMMUNICATIONS
• Volume: 12
• Issue: 1
• Start Page: 4693
• End Page: 4693
• Grant/Funding Information: J.M.B., A.T.B. and R.G. acknowledge NSF GRFP grant no. 1444932. J.M.B. acknowledges NCI fellowship grant no. F99CA234959. A.T.B. acknowledges NCI fellowship grant no. F99CA245789. A.V.K. acknowledges NIH grant no. F31 F31CA243502. M.E.Q. acknowledges NIH grant no. F31 F31HL134241. R.L. acknowledges NIH grant no. HL082808. A.L.M. and K.S. acknowledge NIH grant no. R01GM131099. A.L.M. acknowledges NSF CAREER1832100. K.S. acknowledges NIH grant no. R01GM124472
• Keywords: Multidisciplinary Sciences; DYNAMICS; Science & Technology; 3D ORIENTATION; LIVE; FOCAL ADHESIONS; SINGLE; GENERATED FORCES; DNA; Science & Technology - Other Topics; LOCALIZATION; MOLECULE; TENSION PROBES
• Research Categories: Health Sciences, Oncology

Abstract

Many cellular processes, including cell division, development, and cell migration require spatially and temporally coordinated forces transduced by cell-surface receptors. Nucleic acid-based molecular tension probes allow one to visualize the piconewton (pN) forces applied by these receptors. Building on this technology, we recently developed molecular force microscopy (MFM) which uses fluorescence polarization to map receptor force orientation with diffraction-limited resolution (~250 nm). Here, we show that structured illumination microscopy (SIM), a super-resolution technique, can be used to perform super-resolution MFM. Using SIM-MFM, we generate the highest resolution maps of both the magnitude and orientation of the pN traction forces applied by cells. We apply SIM-MFM to map platelet and fibroblast integrin forces, as well as T cell receptor forces. Using SIM-MFM, we show that platelet traction force alignment occurs on a longer timescale than adhesion. Importantly, SIM-MFM can be implemented on any standard SIM microscope without hardware modifications.