Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy

Wei Chun Tang; Yen Ting Liu; Cheng Han Yeh; Chieh Han Lu; Chiao Hui Tu; Yi Ling Lin; Yu Chun Lin; Tsui Ling Hsu; Liang Gao; Shu Wei Chang; Peilin Chen; Bi Chang Chen

doi:10.1038/s42003-022-03835-6

Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy

Wei Chun Tang
, Yen Ting Liu
, Cheng Han Yeh
, Chieh Han Lu
, Chiao Hui Tu
, Yi Ling Lin
, Yu Chun Lin
, Tsui Ling Hsu
, Liang Gao
, Shu Wei Chang
, Peilin Chen^*
, Bi Chang Chen^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

Lattice 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.

Original language	English
Article number	879
Journal	Communications Biology
Volume	5
Issue number	1
DOIs	https://doi.org/10.1038/s42003-022-03835-6
Publication status	Published - 2022 Dec
Externally published	Yes

ASJC Scopus subject areas

Medicine (miscellaneous)
General Biochemistry,Genetics and Molecular Biology
General Agricultural and Biological Sciences

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@article{6dec1bef13514579a484ec469ab55554,

title = "Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy",

abstract = "Lattice 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.",

author = "Tang, \{Wei Chun\} and Liu, \{Yen Ting\} and Yeh, \{Cheng Han\} and Lu, \{Chieh Han\} and Tu, \{Chiao Hui\} and Lin, \{Yi Ling\} and Lin, \{Yu Chun\} and Hsu, \{Tsui Ling\} and Liang Gao and Chang, \{Shu Wei\} and Peilin Chen and Chen, \{Bi Chang\}",

note = "Publisher Copyright: {\textcopyright} 2022, The Author(s).",

year = "2022",

month = dec,

doi = "10.1038/s42003-022-03835-6",

language = "English",

volume = "5",

journal = "Communications Biology",

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T1 - Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy

AU - Tang, Wei Chun

AU - Liu, Yen Ting

AU - Yeh, Cheng Han

AU - Lu, Chieh Han

AU - Tu, Chiao Hui

AU - Lin, Yi Ling

AU - Lin, Yu Chun

AU - Hsu, Tsui Ling

AU - Gao, Liang

AU - Chang, Shu Wei

AU - Chen, Peilin

AU - Chen, Bi Chang

PY - 2022/12

Y1 - 2022/12

N2 - Lattice 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.

AB - Lattice 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.

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DO - 10.1038/s42003-022-03835-6

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JO - Communications Biology

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Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy

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