Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy
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Published:2022-08-26
Issue:1
Volume:5
Page:
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ISSN:2399-3642
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Container-title:Communications Biology
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language:en
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Short-container-title:Commun Biol
Author:
Tang Wei-ChunORCID, Liu Yen-TingORCID, Yeh Cheng-Han, Lu Chieh-HanORCID, Tu Chiao-Hui, Lin Yi-Ling, Lin Yu-ChunORCID, Hsu Tsui-Ling, Gao Liang, Chang Shu-WeiORCID, Chen PeilinORCID, Chen Bi-ChangORCID
Abstract
AbstractLattice lightsheet microscopy (LLSM) featuring three-dimensional recording is improved to manipulate cellular behavior with subcellular resolution through optogenetic activation (optoLLSM). A position-controllable Bessel beam as a stimulation source is integrated into the LLSM to achieve spatiotemporal photoactivation by changing the spatial light modulator (SLM) patterns. Unlike the point-scanning in a confocal microscope, the lattice beams are capable of wide-field optical sectioning for optogenetic activation along the Bessel beam path.We show that the energy power required for optogenetic activations is lower than 1 nW (or 24 mWcm-2) for time-lapses of CRY2olig clustering proteins, and membrane ruffling can be induced at different locations within a cell with subcellular resolution through light-triggered recruitment of phosphoinositide 3-kinase. Moreover, with the epidermal growth factor receptor (EGFR) fused with CRY2olig, we are able to demonstrate guided cell migration using optogenetic stimulation for up to 6 h, where 463 imaging volumes are collected, without noticeable cellular damages.
Funder
Academia Sinica Ministry of Science and Technology, Taiwan
Publisher
Springer Science and Business Media LLC
Subject
General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology,Medicine (miscellaneous)
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