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Article ; Online: Ciliary localization of a light-activated neuronal GPCR shapes behavior.

Winans, Amy M / Friedmann, Drew / Stanley, Cherise / Xiao, Tong / Liu, Tsung-Li / Chang, Christopher J / Isacoff, Ehud Y

Proceedings of the National Academy of Sciences of the United States of America

2023 Volume 120, Issue 43, Page(s) e2311131120

Abstract: Many neurons in the central nervous system produce a single primary cilium that serves as a specialized signaling organelle. Several neuromodulatory G-protein-coupled receptors (GPCRs) localize to primary cilia in neurons, although it is not understood ... ...

Abstract	Many neurons in the central nervous system produce a single primary cilium that serves as a specialized signaling organelle. Several neuromodulatory G-protein-coupled receptors (GPCRs) localize to primary cilia in neurons, although it is not understood how GPCR signaling from the cilium impacts circuit function and behavior. We find that the vertebrate ancient long opsin A (VALopA), a G
MeSH term(s)	Animals ; Zebrafish ; Receptors, G-Protein-Coupled/physiology ; Signal Transduction/physiology ; Opsins ; Rod Opsins ; Neurons ; Cilia/physiology
Chemical Substances	Receptors, G-Protein-Coupled ; Opsins ; Rod Opsins
Language	English
Publishing date	2023-10-16
Publishing country	United States
Document type	Journal Article
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.2311131120
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Article ; Online: Two-dimensional hybrid photonic/plasmonic crystal cavities.

Liu, Tsung-li / Russell, Kasey J / Cui, Shanying / Hu, Evelyn L

Optics express

2014 Volume 22, Issue 7, Page(s) 8219–8225

Abstract: We present a 2-D plasmonic crystal design with visible band-gap by combining a 2-D photonic crystal with TM band-gap and a silver surface. Simulations show that the presence of the silver surface gives rise to an expanded band-gap. A plasmonic crystal ... ...

Abstract	We present a 2-D plasmonic crystal design with visible band-gap by combining a 2-D photonic crystal with TM band-gap and a silver surface. Simulations show that the presence of the silver surface gives rise to an expanded band-gap. A plasmonic crystal defect cavity with Q ~300 and mode volume ~1.9x10(-2) (λ/n) (3) can be formed using our design. The total Q of such a cavity is determined by both the radiative loss of the dielectric component, as well as absorption loss to the metal. We provide design criteria for the optimization of the total Q to allow high radiative or extraction efficiency.
Language	English
Publishing date	2014-04-07
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1491859-6
ISSN	1094-4087 ; 1094-4087
ISSN (online)	1094-4087
ISSN	1094-4087
DOI	10.1364/OE.22.008219
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Article ; Online: 4D cell biology: big data image analytics and lattice light-sheet imaging reveal dynamics of clathrin-mediated endocytosis in stem cell-derived intestinal organoids.

Schöneberg, Johannes / Dambournet, Daphné / Liu, Tsung-Li / Forster, Ryan / Hockemeyer, Dirk / Betzig, Eric / Drubin, David G

Molecular biology of the cell

2018 Volume 29, Issue 24, Page(s) 2959–2968

Abstract: New methods in stem cell 3D organoid tissue culture, advanced imaging, and big data image analytics now allow tissue-scale 4D cell biology, but currently available analytical pipelines are inadequate for handing and analyzing the resulting gigabytes and ... ...

Abstract	New methods in stem cell 3D organoid tissue culture, advanced imaging, and big data image analytics now allow tissue-scale 4D cell biology, but currently available analytical pipelines are inadequate for handing and analyzing the resulting gigabytes and terabytes of high-content imaging data. We expressed fluorescent protein fusions of clathrin and dynamin2 at endogenous levels in genome-edited human embryonic stem cells, which were differentiated into hESC-derived intestinal epithelial organoids. Lattice light-sheet imaging with adaptive optics (AO-LLSM) allowed us to image large volumes of these organoids (70 × 60 × 40 µm xyz) at 5.7 s/frame. We developed an open-source data analysis package termed pyLattice to process the resulting large (∼60 Gb) movie data sets and to track clathrin-mediated endocytosis (CME) events. CME tracks could be recorded from ∼35 cells at a time, resulting in ∼4000 processed tracks per movie. On the basis of their localization in the organoid, we classified CME tracks into apical, lateral, and basal events and found that CME dynamics is similar for all three classes, despite reported differences in membrane tension. pyLattice coupled with AO-LLSM makes possible quantitative high temporal and spatial resolution analysis of subcellular events within tissues.
MeSH term(s)	Animals ; Big Data ; Cell Culture Techniques/methods ; Cell Differentiation/physiology ; Clathrin/metabolism ; Clathrin-Coated Vesicles/metabolism ; Dynamin II/metabolism ; Endocytosis/physiology ; Human Embryonic Stem Cells/cytology ; Human Embryonic Stem Cells/metabolism ; Humans ; Image Processing, Computer-Assisted/methods ; Intestinal Mucosa/cytology ; Intestinal Mucosa/metabolism ; Intestines/cytology ; Mice ; Organoids/cytology ; Organoids/diagnostic imaging ; Organoids/metabolism
Chemical Substances	Clathrin ; Dynamin II (EC 3.6.5.5)
Language	English
Publishing date	2018-09-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.E18-06-0375
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Zs.A 2853: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 410: Show issues

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Article ; Online: Controlled mode tuning in 1-D 'RIM' plasmonic crystal trench cavities probed with coupled optical emitters.

Liu, Tsung-li / Russell, Kasey J / Cui, Shanying / Hu, Evelyn L

Optics express

2013 Volume 21, Issue 24, Page(s) 30074–30081

Abstract: We present a design of plasmonic cavities that consists of two sets of 1-D plasmonic crystal reflectors on a plasmonic trench waveguide. A 'reverse image mold' (RIM) technique was developed to pattern high-resolution silver trenches and to embed emitters ...

Abstract	We present a design of plasmonic cavities that consists of two sets of 1-D plasmonic crystal reflectors on a plasmonic trench waveguide. A 'reverse image mold' (RIM) technique was developed to pattern high-resolution silver trenches and to embed emitters at the cavity field maximum, and FDTD simulations were performed to analyze the frequency response of the fabricated devices. Distinct cavity modes were observed from the photoluminescence spectra of the organic dye embedded within these cavities. The cavity geometry facilitates tuning of the modes through a change in cavity dimensions. Both the design and the fabrication technique presented could be extended to making trench waveguide-based plasmonic devices and circuits.
Language	English
Publishing date	2013-12-02
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1491859-6
ISSN	1094-4087 ; 1094-4087
ISSN (online)	1094-4087
ISSN	1094-4087
DOI	10.1364/OE.21.030074
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Article ; Online: Visualizing dynamic microvillar search and stabilization during ligand detection by T cells.

Cai, En / Marchuk, Kyle / Beemiller, Peter / Beppler, Casey / Rubashkin, Matthew G / Weaver, Valerie M / Gérard, Audrey / Liu, Tsung-Li / Chen, Bi-Chang / Betzig, Eric / Bartumeus, Frederic / Krummel, Matthew F

Science (New York, N.Y.)

2017 Volume 356, Issue 6338

Abstract: During immune surveillance, T cells survey the surface of antigen-presenting cells. In searching for peptide-loaded major histocompatibility complexes (pMHCs), they must solve a classic trade-off between speed and sensitivity. It has long been supposed ... ...

Abstract	During immune surveillance, T cells survey the surface of antigen-presenting cells. In searching for peptide-loaded major histocompatibility complexes (pMHCs), they must solve a classic trade-off between speed and sensitivity. It has long been supposed that microvilli on T cells act as sensory organs to enable search, but their strategy has been unknown. We used lattice light-sheet and quantum dot-enabled synaptic contact mapping microscopy to show that anomalous diffusion and fractal organization of microvilli survey the majority of opposing surfaces within 1 minute. Individual dwell times were long enough to discriminate pMHC half-lives and T cell receptor (TCR) accumulation selectively stabilized microvilli. Stabilization was independent of tyrosine kinase signaling and the actin cytoskeleton, suggesting selection for avid TCR microclusters. This work defines the efficient cellular search process against which ligand detection takes place.
MeSH term(s)	Actin Cytoskeleton/metabolism ; Animals ; Antigens/immunology ; Fractals ; Ligands ; Mice ; Microscopy/methods ; Microvilli/chemistry ; Microvilli/metabolism ; Quantum Dots ; Receptors, Antigen, T-Cell/metabolism ; T-Lymphocytes/cytology ; T-Lymphocytes/immunology ; T-Lymphocytes/metabolism
Chemical Substances	Antigens ; Ligands ; Receptors, Antigen, T-Cell
Language	English
Publishing date	2017-05-11
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	128410-1
ISSN	1095-9203 ; 0036-8075
ISSN (online)	1095-9203
ISSN	0036-8075
DOI	10.1126/science.aal3118
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Zs.A 27: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 77: Show issues

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Article ; Online: Lamellar projections in the endolymphatic sac act as a relief valve to regulate inner ear pressure.

Swinburne, Ian A / Mosaliganti, Kishore R / Upadhyayula, Srigokul / Liu, Tsung-Li / Hildebrand, David G C / Tsai, Tony Y-C / Chen, Anzhi / Al-Obeidi, Ebaa / Fass, Anna K / Malhotra, Samir / Engert, Florian / Lichtman, Jeff W / Kirchhausen, Tomas / Betzig, Eric / Megason, Sean G

eLife

2018 Volume 7

Abstract: The inner ear is a fluid-filled closed-epithelial structure whose function requires maintenance of an internal hydrostatic pressure and fluid composition. The endolymphatic sac (ES) is a dead-end epithelial tube connected to the inner ear whose function ... ...

Abstract	The inner ear is a fluid-filled closed-epithelial structure whose function requires maintenance of an internal hydrostatic pressure and fluid composition. The endolymphatic sac (ES) is a dead-end epithelial tube connected to the inner ear whose function is unclear. ES defects can cause distended ear tissue, a pathology often seen in hearing and balance disorders. Using live imaging of zebrafish larvae, we reveal that the ES undergoes cycles of slow pressure-driven inflation followed by rapid deflation. Absence of these cycles in
MeSH term(s)	Animals ; Animals, Genetically Modified ; Embryo, Nonmammalian ; Endolymphatic Sac/anatomy & histology ; Endolymphatic Sac/physiology ; Endolymphatic Sac/ultrastructure ; Female ; Gene Expression ; Hearing/physiology ; Homeostasis/physiology ; Hydrostatic Pressure ; In Situ Hybridization, Fluorescence ; Larva/anatomy & histology ; Larva/physiology ; Larva/ultrastructure ; Male ; Microscopy, Electron ; Mutation ; Time-Lapse Imaging ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Zebrafish ; Zebrafish Proteins/genetics ; Zebrafish Proteins/metabolism
Chemical Substances	Transcription Factors ; Zebrafish Proteins ; lmx1bb protein, zebrafish
Language	English
Publishing date	2018-06-19
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.37131
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Article ; Online: High-resolution live imaging reveals axon-glia interactions during peripheral nerve injury and repair in zebrafish.

Xiao, Yan / Faucherre, Adèle / Pola-Morell, Laura / Heddleston, John M / Liu, Tsung-Li / Chew, Teng-Leong / Sato, Fuminori / Sehara-Fujisawa, Atsuko / Kawakami, Koichi / López-Schier, Hernán

Disease models & mechanisms

2015 Volume 8, Issue 6, Page(s) 553–564

Abstract: Neural damage is a devastating outcome of physical trauma. The glia are one of the main effectors of neuronal repair in the nervous system, but the dynamic interactions between peripheral neurons and Schwann cells during injury and regeneration remain ... ...

Abstract	Neural damage is a devastating outcome of physical trauma. The glia are one of the main effectors of neuronal repair in the nervous system, but the dynamic interactions between peripheral neurons and Schwann cells during injury and regeneration remain incompletely characterized. Here, we combine laser microsurgery, genetic analysis, high-resolution intravital imaging and lattice light-sheet microscopy to study the interaction between Schwann cells and sensory neurons in a zebrafish model of neurotrauma. We found that chronic denervation by neuronal ablation leads to Schwann-cell death, whereas acute denervation by axonal severing does not affect the overall complexity and architecture of the glia. Neuronal-circuit regeneration begins when Schwann cells extend bridging processes to close the injury gap. Regenerating axons grow faster and directionally after the physiological clearing of distal debris by the Schwann cells. This might facilitate circuit repair by ensuring that axons are guided through unoccupied spaces within bands of Büngner towards their original peripheral target. Accordingly, in the absence of Schwann cells, regenerating axons are misrouted, impairing the re-innervation of sensory organs. Our results indicate that regenerating axons use haptotaxis as a directional cue during the reconstitution of a neural circuit. These findings have implications for therapies aimed at neurorepair, which will benefit from preserving the architecture of the peripheral glia during periods of denervation.
MeSH term(s)	Animals ; Animals, Genetically Modified ; Axons/metabolism ; Cell Communication ; Chronic Disease ; Claudins/metabolism ; Denervation ; Homozygote ; Lasers ; Molecular Imaging/methods ; Mutagenesis, Insertional/genetics ; Myelin-Associated Glycoprotein/metabolism ; Nerve Regeneration ; Neuroglia/metabolism ; Peripheral Nerve Injuries/pathology ; Phenotype ; Schwann Cells/metabolism ; Zebrafish/physiology ; Zebrafish Proteins/genetics
Chemical Substances	Claudins ; Myelin-Associated Glycoprotein ; Zebrafish Proteins
Language	English
Publishing date	2015-06
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1754-8411
ISSN (online)	1754-8411
DOI	10.1242/dmm.018184
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Article ; Online: Membrane dynamics of dividing cells imaged by lattice light-sheet microscopy.

Aguet, François / Upadhyayula, Srigokul / Gaudin, Raphaël / Chou, Yi-Ying / Cocucci, Emanuele / He, Kangmin / Chen, Bi-Chang / Mosaliganti, Kishore / Pasham, Mithun / Skillern, Wesley / Legant, Wesley R / Liu, Tsung-Li / Findlay, Greg / Marino, Eric / Danuser, Gaudenz / Megason, Sean / Betzig, Eric / Kirchhausen, Tom

Molecular biology of the cell

2016 Volume 27, Issue 22, Page(s) 3418–3435

Abstract: Membrane remodeling is an essential part of transferring components to and from the cell surface and membrane-bound organelles and for changes in cell shape, which are particularly critical during cell division. Earlier analyses, based on classical ... ...

Abstract	Membrane remodeling is an essential part of transferring components to and from the cell surface and membrane-bound organelles and for changes in cell shape, which are particularly critical during cell division. Earlier analyses, based on classical optical live-cell imaging and mostly restricted by technical necessity to the attached bottom surface, showed persistent formation of endocytic clathrin pits and vesicles during mitosis. Taking advantage of the resolution, speed, and noninvasive illumination of the newly developed lattice light-sheet fluorescence microscope, we reexamined their assembly dynamics over the entire cell surface and found that clathrin pits form at a lower rate during late mitosis. Full-cell imaging measurements of cell surface area and volume throughout the cell cycle of single cells in culture and in zebrafish embryos showed that the total surface increased rapidly during the transition from telophase to cytokinesis, whereas cell volume increased slightly in metaphase and was relatively constant during cytokinesis. These applications demonstrate the advantage of lattice light-sheet microscopy and enable a new standard for imaging membrane dynamics in single cells and multicellular assemblies.
MeSH term(s)	Animals ; Cell Cycle ; Cell Membrane/physiology ; Cell Membrane/ultrastructure ; Clathrin/metabolism ; Cytokinesis/physiology ; Endosomes/metabolism ; Imaging, Three-Dimensional/methods ; Metaphase ; Microscopy/methods ; Microscopy, Fluorescence/methods ; Mitosis/physiology ; Zebrafish/embryology
Chemical Substances	Clathrin
Language	English
Publishing date	2016-08-17
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.E16-03-0164
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Zs.A 2853: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 410: Show issues

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Article ; Online: Cytoskeletal actin dynamics shape a ramifying actin network underpinning immunological synapse formation.

Fritzsche, Marco / Fernandes, Ricardo A / Chang, Veronica T / Colin-York, Huw / Clausen, Mathias P / Felce, James H / Galiani, Silvia / Erlenkämper, Christoph / Santos, Ana M / Heddleston, John M / Pedroza-Pacheco, Isabela / Waithe, Dominic / de la Serna, Jorge Bernardino / Lagerholm, B Christoffer / Liu, Tsung-Li / Chew, Teng-Leong / Betzig, Eric / Davis, Simon J / Eggeling, Christian

Science advances

2017 Volume 3, Issue 6, Page(s) e1603032

Abstract: T cell activation and especially trafficking of T cell receptor microclusters during immunological synapse formation are widely thought to rely on cytoskeletal remodeling. However, important details on the involvement of actin in the latter transport ... ...

Abstract	T cell activation and especially trafficking of T cell receptor microclusters during immunological synapse formation are widely thought to rely on cytoskeletal remodeling. However, important details on the involvement of actin in the latter transport processes are missing. Using a suite of advanced optical microscopes to analyze resting and activated T cells, we show that, following contact formation with activating surfaces, these cells sequentially rearrange their cortical actin across the entire cell, creating a previously unreported ramifying actin network above the immunological synapse. This network shows all the characteristics of an inward-growing transportation network and its dynamics correlating with T cell receptor rearrangements. This actin reorganization is accompanied by an increase in the nanoscale actin meshwork size and the dynamic adjustment of the turnover times and filament lengths of two differently sized filamentous actin populations, wherein formin-mediated long actin filaments support a very flat and stiff contact at the immunological synapse interface. The initiation of immunological synapse formation, as highlighted by calcium release, requires markedly little contact with activating surfaces and no cytoskeletal rearrangements. Our work suggests that incipient signaling in T cells initiates global cytoskeletal rearrangements across the whole cell, including a stiffening process for possibly mechanically supporting contact formation at the immunological synapse interface as well as a central ramified transportation network apparently directed at the consolidation of the contact and the delivery of effector functions.
MeSH term(s)	Actin Cytoskeleton/metabolism ; Actins/metabolism ; Animals ; Biomarkers ; Cell Line ; Cytoskeleton ; Gene Rearrangement, T-Lymphocyte ; Humans ; Immunological Synapses/metabolism ; Lymphocyte Activation ; Receptors, Antigen, T-Cell/genetics ; Receptors, Antigen, T-Cell/metabolism ; Signal Transduction ; T-Lymphocytes/immunology ; T-Lymphocytes/metabolism
Chemical Substances	Actins ; Biomarkers ; Receptors, Antigen, T-Cell
Language	English
Publishing date	2017-06-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2810933-8
ISSN	2375-2548 ; 2375-2548
ISSN (online)	2375-2548
ISSN	2375-2548
DOI	10.1126/sciadv.1603032
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Article ; Online: Cortical column and whole-brain imaging with molecular contrast and nanoscale resolution.

Gao, Ruixuan / Asano, Shoh M / Upadhyayula, Srigokul / Pisarev, Igor / Milkie, Daniel E / Liu, Tsung-Li / Singh, Ved / Graves, Austin / Huynh, Grace H / Zhao, Yongxin / Bogovic, John / Colonell, Jennifer / Ott, Carolyn M / Zugates, Christopher / Tappan, Susan / Rodriguez, Alfredo / Mosaliganti, Kishore R / Sheu, Shu-Hsien / Pasolli, H Amalia /

Pang, Song / Xu, C Shan / Megason, Sean G / Hess, Harald / Lippincott-Schwartz, Jennifer / Hantman, Adam / Rubin, Gerald M / Kirchhausen, Tom / Saalfeld, Stephan / Aso, Yoshinori / Boyden, Edward S / Betzig, Eric

Science (New York, N.Y.)

2019 Volume 363, Issue 6424

Abstract: Optical and electron microscopy have made tremendous inroads toward understanding the complexity of the brain. However, optical microscopy offers insufficient resolution to reveal subcellular details, and electron microscopy lacks the throughput and ... ...

Abstract	Optical and electron microscopy have made tremendous inroads toward understanding the complexity of the brain. However, optical microscopy offers insufficient resolution to reveal subcellular details, and electron microscopy lacks the throughput and molecular contrast to visualize specific molecular constituents over millimeter-scale or larger dimensions. We combined expansion microscopy and lattice light-sheet microscopy to image the nanoscale spatial relationships between proteins across the thickness of the mouse cortex or the entire
MeSH term(s)	Animals ; Axons ; Brain/diagnostic imaging ; Dendritic Spines ; Drosophila ; Female ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Kidney/diagnostic imaging ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microscopy, Fluorescence ; Nanotechnology ; Neuroimaging/methods ; Optical Imaging/methods ; Phantoms, Imaging ; Somatosensory Cortex/diagnostic imaging ; Synapses
Language	English
Publishing date	2019-01-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Video-Audio Media
ZDB-ID	128410-1
ISSN	1095-9203 ; 0036-8075
ISSN (online)	1095-9203
ISSN	0036-8075
DOI	10.1126/science.aau8302
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