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

2 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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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).",

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month = dec,

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

language = "English",

volume = "5",

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

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