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bioRxiv · 10.1101/2022.01.02.474058

Optogenetic Manipulation of Cell Migration with High Spatiotemporal Resolution Using Lattice Lightsheet Microscopy

Tang, W.-C.·Liu, Y.-T.·Yeh, C.-H.·Lin, Y.-L.·Lin, Y.-C.·Hsu, T.-L.·Gao, L.·Chang, S.-W.·Chen, P.·Chen, B.-C.
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Abstract

Lattice lightsheet microscopy (LLSM) is modified with the aim of manipulating cellular behavior with subcellular resolution through three-dimensional (3D) optogenetic activation. In this study, we report a straightforward implementation of the activation source in LLSM in which the stimulating light can be generated by changing the spatial light modulator (SLM) patterns and the annual masks. As a result, a Bessel beam as a stimulation source is integrated into the LLSM without changing the optical configuration, achieving better spatiotemporal control of photoactivation. We show that the energy power required for optogenetic reactions is lower than 1 nW (or 24 mW/cm2), and membrane ruffling can be activated at different locations within a cell with subcellular resolution. We also demonstrate guided cell migration using optogenetic stimulation for up to 6 h, where 463 imaging volumes are collected, without noticeable damage to cells.

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Tang, W.-C., Liu, Y.-T., Yeh, C.-H., Lin, Y.-L., Lin, Y.-C., Hsu, T.-L., Gao, L., Chang, S.-W., Chen, P., Chen, B.-C.. 2022-01-02. Optogenetic Manipulation of Cell Migration with High Spatiotemporal Resolution Using Lattice Lightsheet Microscopy. https://doi.org/10.1101/2022.01.02.474058

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