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Superresolved microparticle traction force microscopy reveals subcellular force patterns in immune cell-target interactions

Daan Vorselen, Yifan Wang, Miguel M. de Jesus, Pavak K. Shah, Matthew J. Footer, Morgan Huse, Wei Cai, Julie A. Theriot
doi: https://doi.org/10.1101/431221
Daan Vorselen
1Department of Biochemistry, Stanford University, Stanford, CA 94305, USA
2Department of Biology and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98105, USA
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Yifan Wang
3Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, USA
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Miguel M. de Jesus
4Immunology Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
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Pavak K. Shah
5Developmental Biology Program, Sloan Kettering Institute, New York, NY 10065, USA
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Matthew J. Footer
1Department of Biochemistry, Stanford University, Stanford, CA 94305, USA
2Department of Biology and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98105, USA
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Morgan Huse
4Immunology Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
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Wei Cai
3Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, USA
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Julie A. Theriot
1Department of Biochemistry, Stanford University, Stanford, CA 94305, USA
2Department of Biology and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98105, USA
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  • For correspondence: jtheriot@uw.edu
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Abstract

Force exertion is an integral part of cellular behavior. Traction force microscopy (TFM) has been instrumental for studying such forces, providing both spatial and directional force measurements at subcellular resolution. However, the applications of classical TFM are restricted by the typical planar geometry. Here, we develop a particle-based force sensing strategy, specifically designed for studying ligand-dependent cellular interactions. We establish a straightforward batch approach for synthesizing highly uniform, deformable and tunable hydrogel particles, which can also be easily derivatized to trigger specific cellular behavior. The 3D shape of such particles can be resolved with superresolution (<50 nm) accuracy using conventional confocal microscopy. We introduce a computational method that allows inference of surface traction forces with high sensitivity (∼10 Pa) directly from the particle shape. We illustrate the potential and flexibility of this approach by revealing surprising subcellular force patterns throughout phagocytic engulfment and measuring dynamics of cytotoxic T cell force exertion in the immunological synapse. This strategy can readily be adapted for studying cellular forces in a wide range of applications.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted March 13, 2019.
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Superresolved microparticle traction force microscopy reveals subcellular force patterns in immune cell-target interactions
Daan Vorselen, Yifan Wang, Miguel M. de Jesus, Pavak K. Shah, Matthew J. Footer, Morgan Huse, Wei Cai, Julie A. Theriot
bioRxiv 431221; doi: https://doi.org/10.1101/431221
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Superresolved microparticle traction force microscopy reveals subcellular force patterns in immune cell-target interactions
Daan Vorselen, Yifan Wang, Miguel M. de Jesus, Pavak K. Shah, Matthew J. Footer, Morgan Huse, Wei Cai, Julie A. Theriot
bioRxiv 431221; doi: https://doi.org/10.1101/431221

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