Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy
Weike Tang,Yen-Ting Liu,Cheng-Han Yeh,Yi-Ling Lin,Yu-Cheng Lin,Tsui-Ling Hsu,Liang Gao,Shu-Wei Chang,Peilin Chen,Bi-Chang Chen +9 more
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In this article , a position-controllable Bessel beam is integrated into the LLSM to achieve spatiotemporal photoactivation by changing the spatial light modulator (SLM) patterns.Abstract:
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. read more
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References
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Collective cell migration in morphogenesis, regeneration and cancer
Peter Friedl,Darren Gilmour +1 more
TL;DR: Comparing different types of collective migration at the molecular and cellular level reveals a common mechanistic theme between developmental and cancer research.
Journal ArticleDOI
Lattice Light Sheet Microscopy: Imaging Molecules to Embryos at High Spatiotemporal Resolution
Bi-Chang Chen,Wesley R. Legant,Kai Wang,Lin Shao,Daniel E. Milkie,Michael W. Davidson,Chris Janetopoulos,Xufeng S. Wu,John A. Hammer,Zhe Liu,Brian P. English,Yuko Mimori-Kiyosue,Daniel P. Romero,Alex T. Ritter,Alex T. Ritter,Jennifer Lippincott-Schwartz,Lillian K. Fritz-Laylin,R. Dyche Mullins,Diana M. Mitchell,Joshua N. Bembenek,Anne-Cécile Reymann,Ralph Böhme,Stephan W. Grill,Jennifer T. Wang,Geraldine Seydoux,U. Serdar Tulu,Daniel P. Kiehart,Eric Betzig +27 more
TL;DR: A new microscope using ultrathin light sheets derived from two-dimensional optical lattices is developed, demonstrating the performance advantages of lattice light-sheet microscopy compared with previous techniques and highlighted phenomena that, when seen at increased spatiotemporal detail, may hint at previously unknown biological mechanisms.
Journal ArticleDOI
Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination
Thomas A. Planchon,Liang Gao,Daniel E. Milkie,Michael W. Davidson,James A. Galbraith,Catherine G. Galbraith,Eric Betzig +6 more
TL;DR: Scanned Bessel beams are used in conjunction with structured illumination and/or two-photon excitation to create thinner light sheets better suited to three-dimensional (3D) subcellular imaging.
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Rapid blue-light-mediated induction of protein interactions in living cells
Matthew J. Kennedy,Robert M. Hughes,Leslie A. Peteya,Joel W. Schwartz,Michael D. Ehlers,Michael D. Ehlers,Chandra L. Tucker +6 more
TL;DR: Genetically encoded light-inducible protein-interaction modules based on Arabidopsis thaliana cryptochrome 2 and CIB1 that require no exogenous ligands and dimerize on blue-light exposure with subsecond time resolution and subcellular spatial resolution are described.
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Cdc42 - the centre of polarity
TL;DR: It is now clear that many features of the molecular mechanisms controlling polarization are conserved in all eukaryotic cells, including Cdc42, a small GTPase of the Rho family.
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