51. Quantitative biophysical methods to study immune cell mechanobiology
- Author
-
Barbieri, Liliana, Fritzsche, Marco, Colin York, Huw Ewryd, and Eggeling, Christian
- Subjects
Immunology ,Microscopy ,Biophysics - Abstract
Mechanobiology is increasingly thought to critically underpin health and disease. During their lifetime, cells carry out their function interacting with the external biophysical environment through mechanical and biochemical events. This is the case for T cells of the adaptive immune system during their activation via interactions with Antigen Presenting Cells (APCs). T-cell activation critically depends on the binding of the T-cell receptor (TCR) to the foreign antigen presented by the APC at the immunological synapse, leading to an orchestrated series of events including cytoskeletal rearrangement, the release of calcium from the endoplasmic reticulum and a change in the expression of surface proteins. TCR cluster migration is a consequence of actin cytoskeleton mediated forces in the range of pico-Newton and sub-second time scale. Hence, dissecting the significance of mechanobiology during T-cell activation requires quantitative biophysical methods for force and calcium readouts. However, the existing technologies do not offer the required sensitivity, robust quantification, and high statistical power. In this work, we present three new methods for planar and axial force generation on the nanometre length scale and sub-second time scale, combining Traction Force Microscopy with Structured Illumination Microscopy. Next, we establish a new machine learning based single-cell high-throughput platform, to quantify automatically and simultaneously the calcium responses of > 100,000 cells. Taken together, these techniques represent a powerful quantitative biophysical platform, opening up new directions for interrogating the role of mechanobiology in health and diseases.
- Published
- 2021