Turn-key mapping of cell receptor force orientation and magnitude using a commercial structured illumination microscope
| Autor: | Wenchun Chen, Alisina Bazrafshan, Anna V. Kellner, Rachel L. Bender, Khalid Salaita, Roxanne Glazier, Joshua M. Brockman, J. Dale Combs, Hanquan Su, Aaron T. Blanchard, M. Edward Quach, Alexa L. Mattheyses, Renhao Li |
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| Rok vydání: | 2021 |
| Předmět: |
Blood Platelets
Integrins Microscope Materials science Science Traction (engineering) Receptors Antigen T-Cell General Physics and Astronomy Receptors Cell Surface CD8-Positive T-Lymphocytes Time-Lapse Imaging Article General Biochemistry Genetics and Molecular Biology law.invention Mice law Microscopy Animals Humans Super-resolution microscopy Author Correction Fluorescent Dyes Nanoscale biophysics Multidisciplinary Tractive force Tension (physics) Orientation (computer vision) Resolution (electron density) General Chemistry Adhesion Biomechanical Phenomena Biosensors Microscopy Fluorescence Polarization microscopy Molecular Probes NIH 3T3 Cells Paxillin Biological system |
| Zdroj: | Nature Communications Nature Communications, Vol 12, Iss 1, Pp 1-15 (2021) |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/s41467-021-24602-x |
| Popis: | 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. The authors have recently developed molecular force microscopy (MFM) which uses fluorescence polarisation to measure cell-surface receptor force orientation. Here they show that structured illumination microscopes, which inherently use fluorescence polarisation, can be used for MFM in a turn-key manner. |
| Databáze: | OpenAIRE |
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