We focus on the development of optical tools to help study the cell molecular processes in vivo, specially those connected to membrane trafficking and the activity of cytoskeletal molecular motors. These processes are central to sustaining many important cellular functions, such as vesicle transport or cell division, and are therefore being thoroughly studied in vitro by use of a host of new technologies.Despite their success, in vitro experiments can hardly recreate the extraordinary complexity and richness of the cellular interior at the molecular scale. Thus, mechanochemicaldata obtained in vitro will eventually have to be completed and validated against results obtained within living samples, which are unfortunately still very much scarce.
We hope to contribute bridging the gap by adapting existing optical tools to this endeavour in two main directions: real-time manipulation of samples with holographic optical tweezers and development of compatible force-measurement methods valid inside the cytoplasm of living cells.Our approach to measure forces based on a direct estimation of light momentum changes may provide reliable information about cell inner processes, such as the forces exerted by molecular motors during their journey through the intricately-woven, filamentous networks of the cytoskeleton.
Holographic optical traps
Studies in living cells
Algorithms: Design of algorithms to compute holograms for interactive manipulation.
Assessment of the induced laser heating effects on NG108 cells by studying the induction of apoptosis and necrosis on these cells.
Stability of SLMs: Analysis of the temporal instabilities of holographic traps for analog- and digitally-addressed spatial light modulators.
Manipulation of protein bodies: Evaluation of the use of artificially-induced protein bodies as optimized handles for optical trapping experiments.
Holographic dumbbell: Highly-stable dual-trap design using digital holography.
Motor proteins stepping: High-speed tracking with nanometer accuracy of motor-driven structures in living cells.
Direct force measurement
Automatic trapping: Development of a software for automatic trapping of moving targets.
Direct force method: Adaptation of the measurement of light momentum changes to single-beam optical traps.
Measuring forces inside cells: Application of the method based on the detection of light momentum changes to measure forces of molecular motors in vivo.