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

Autor: Blanchard A; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA., Combs JD; Department of Chemistry, Emory University, Atlanta, GA, USA., Brockman JM; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA., Kellner AV; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA., Glazier R; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA., Su H; Department of Chemistry, Emory University, Atlanta, GA, USA., Bender RL; Department of Chemistry, Emory University, Atlanta, GA, USA., Bazrafshan AS; Department of Chemistry, Emory University, Atlanta, GA, USA., Chen W; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Atlanta, GA, USA.; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA., Quach ME; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Atlanta, GA, USA.; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA., Li R; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Atlanta, GA, USA.; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA., Mattheyses AL; Department of Cell, Developmental, and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, USA., Salaita K; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA. k.salaita@emory.edu.; Department of Chemistry, Emory University, Atlanta, GA, USA. k.salaita@emory.edu.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2021 Aug 03; Vol. 12 (1), pp. 4693. Date of Electronic Publication: 2021 Aug 03.
DOI: 10.1038/s41467-021-24602-x
Abstrakt: 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.
(© 2021. The Author(s).)
Databáze: MEDLINE
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