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BP: Fachverband Biologische Physik

BP 8: Cytoskeletal filaments

BP 8.3: Vortrag

Dienstag, 2. April 2019, 10:00–10:15, H10

Studying Cytoskeletal Processes, including T-cell activation, at the single-molecule level with optical tweezers and correlated fluorescence microscopy (OT-CFM) — •Ann Mukhortava, Aida Llauró Portell, Roleand van Wijk, Andrea Candelli, and Gerrit Sitters — LUMICKS, Amsterdam, Netherlands

Microtubules, actin and intermediate filaments are highly dynamic cytoskeletal structures that interact with various motor proteins and regulatory factors and thus play fundamental roles in many essential biological processes. A lot of these, including cell division, signaling, and migration, involve mechano-chemical and -biological pathways. Force spectroscopy on a single-molecule and single-cell level permits exploring and manipulating these complex interactions to help understand their nature better. OT-CFM which we commercialized as the C-Trap™ integrates optical tweezers, confocal/STED microscopy, and an advanced microfluidics system in a truly correlated manner. It enables live, simultaneous and correlative visualization and manipulation of molecular interactions with sub-picoNewton (pN) force resolution and microsecond temporal resolution. Here, we present our experiments on visualizing and quantifying the elastic properties of filaments, the motility of cytoskeletal molecular motors and the force-triggered activation of T-cells using the C-Trap™ system. These experiments show that technological advances in hybrid single-molecule methods can be turned into an easy-to-use and stable instrument that opens up new venues in many research areas.