The Experts below are selected from a list of 9528 Experts worldwide ranked by ideXlab platform
Hideaki Kano - One of the best experts on this subject based on the ideXlab platform.
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multimodal nonlinear optical imaging of caenorhabditis elegans with multiplex coherent anti stokes raman scattering third harmonic generation second harmonic generation and two photon excitation fluorescence
Applied Physics Express, 2020Co-Authors: Shinichi Miyazaki, Philippe Leproux, Vincent Couderc, Yu Hayashi, Hideaki KanoAbstract:Multimodal nonlinear optical microscopy was applied to the nematode Caenorhabditis elegans (C. elegans). Transgenic worms expressing the Yellow Fluorescent Protein (YFP) variant Venus underwent analyses. The lipid and Protein distribution was clearly visualized using multiplex coherent anti-Stokes Raman scattering (CARS), second-harmonic generation (SHG), third-harmonic generation (THG), and two-photon excitation fluorescence (TPEF). The TPEF and SHG signals were found to overlap, which allowed the identification of the ventral nerve cord. These findings demonstrate the powerful potential of multimodal nonlinear optical microscopy to reveal C. elegans microstructures in vivo.
Yu Hayashi - One of the best experts on this subject based on the ideXlab platform.
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multimodal nonlinear optical imaging of caenorhabditis elegans with multiplex coherent anti stokes raman scattering third harmonic generation second harmonic generation and two photon excitation fluorescence
Applied Physics Express, 2020Co-Authors: Shinichi Miyazaki, Philippe Leproux, Vincent Couderc, Yu Hayashi, Hideaki KanoAbstract:Multimodal nonlinear optical microscopy was applied to the nematode Caenorhabditis elegans (C. elegans). Transgenic worms expressing the Yellow Fluorescent Protein (YFP) variant Venus underwent analyses. The lipid and Protein distribution was clearly visualized using multiplex coherent anti-Stokes Raman scattering (CARS), second-harmonic generation (SHG), third-harmonic generation (THG), and two-photon excitation fluorescence (TPEF). The TPEF and SHG signals were found to overlap, which allowed the identification of the ventral nerve cord. These findings demonstrate the powerful potential of multimodal nonlinear optical microscopy to reveal C. elegans microstructures in vivo.
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Multimodal nonlinear optical imaging of Caenorhabditis elegans with multiplex coherent anti-Stokes Raman scattering, third-harmonic generation, second-harmonic generation, and two-photon excitation fluorescence
'IOP Publishing', 2020Co-Authors: Miyazaki Shinichi, Yu Hayashi, Leproux Philippe, Couderc Vincent, Kano HideakiAbstract:International audienceMultimodal nonlinear optical microscopy was used to analyze the nematode Caenorhabditis elegans (C. elegans) expressing Yellow Fluorescent Protein (YFP) in the neurons. The lipid and Protein distributions were clearly visualized using multiplex coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), third harmonic generation (THG), and two-photon excitation fluorescence (TPEF). The TPEF and SHG signals were found to overlap, which allowed identification of the ventral nerve cord. These findings demonstrate the powerful potential of multimodal nonlinear optical microscopy to reveal C. elegans microstructures in vivo
Shinichi Miyazaki - One of the best experts on this subject based on the ideXlab platform.
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multimodal nonlinear optical imaging of caenorhabditis elegans with multiplex coherent anti stokes raman scattering third harmonic generation second harmonic generation and two photon excitation fluorescence
Applied Physics Express, 2020Co-Authors: Shinichi Miyazaki, Philippe Leproux, Vincent Couderc, Yu Hayashi, Hideaki KanoAbstract:Multimodal nonlinear optical microscopy was applied to the nematode Caenorhabditis elegans (C. elegans). Transgenic worms expressing the Yellow Fluorescent Protein (YFP) variant Venus underwent analyses. The lipid and Protein distribution was clearly visualized using multiplex coherent anti-Stokes Raman scattering (CARS), second-harmonic generation (SHG), third-harmonic generation (THG), and two-photon excitation fluorescence (TPEF). The TPEF and SHG signals were found to overlap, which allowed the identification of the ventral nerve cord. These findings demonstrate the powerful potential of multimodal nonlinear optical microscopy to reveal C. elegans microstructures in vivo.
Leo M Chalupa - One of the best experts on this subject based on the ideXlab platform.
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morphological properties of mouse retinal ganglion cells
Neuroscience, 2006Co-Authors: J Coombs, D Van Der List, Guo Yong Wang, Leo M ChalupaAbstract:The mouse retina offers an increasingly valuable model for vision research given the possibilities for genetic manipulation. Here we assess how the structural properties of mouse retinal ganglion cells relate to the stratification pattern of the dendrites of these neurons within the inner plexiform layer. For this purpose, we used 14 morphological measures to classify mouse retinal ganglion cells parametrically into different clusters. Retinal ganglion cells were labeled in one of three ways: Lucifer Yellow injection, 'DiOlistics' or transgenic expression of Yellow Fluorescent Protein. The resulting analysis of 182 cells revealed 10 clusters of monostratified cells, with dendrites confined to either On or Off sublaminae of the inner plexiform layer, and four clusters of bistratified cells, dendrites spanning the On and Off sublaminae. We also sought to establish how these parametrically identified retinal ganglion cell clusters relate to cell types identified previously on the basis of immunocytochemical staining and the expression of Yellow Fluorescent Protein. Cells labeled with an antibody against melanopsin were found to be located within a single cluster, while those labeled with the SMI-32 antibody were in four different clusters. Yellow Fluorescent Protein expressing cells were distributed within 13 of the 14 clusters identified here, which demonstrates that Yellow Fluorescent Protein expression is a useful method for labeling virtually the entire population of mouse retinal ganglion cells. Collectively, these findings provide a valuable baseline for future studies dealing with the effects of genetic mutations on the morphological development of these neurons.
Roger Y. Tsien - One of the best experts on this subject based on the ideXlab platform.
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Detection of calcium transients in Drosophila mushroom body neurons with camgaroo reporters.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 2003Co-Authors: Geoffrey S. Baird, Roger Y. Tsien, Ronald L DavisAbstract:Camgaroos are Yellow Fluorescent Protein derivatives that hold promise as transgenically encoded calcium sensors in behaving animals. We expressed two versions of camgaroo in Drosophila mushroom bodies using the galactosidase-4 (GAL4) system. Potassium depolarization of brains expressing the reporters produces a robust increase in fluorescence that is blocked by removing extracellular calcium or by antagonists of voltage-dependent calcium channels. The fluorescence increase is not attributable to cytoplasmic alkalization; depolarization induces a slight acidification of the cytoplasm of mushroom body neurons. Acetylcholine applied near the dendrites of the mushroom body neurons induces a rapid and ipsilateral-specific fluorescence increase in the mushroom body axons that is blocked by antagonists of calcium channels or nicotinic acetylcholine receptors. Fluorescence was observed to increase in all three classes of mushroom body neurons, indicating that all types respond to cholinergic innervation.
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reducing the environmental sensitivity of Yellow Fluorescent Protein mechanism and applications
Journal of Biological Chemistry, 2001Co-Authors: Oliver Griesbeck, Robert E. Campbell, Geoffrey S Baird, David A Zacharias, Roger Y. TsienAbstract:Yellow mutants of the green Fluorescent Protein (YFP) are crucial constituents of genetically encoded indicators of signal transduction and fusions to monitor ProteinProtein interactions. However, previous YFPs show excessive pH sensitivity, chloride interference, poor photostability, or poor expression at 37 °C. Protein evolution in Escherichia coli has produced a new YFP named Citrine, in which the mutation Q69M confers a much lower pKa (5.7) than for previous YFPs, indifference to chloride, twice the photostability of previous YFPs, and much better expression at 37 °C and in organelles. The halide resistance is explained by a 2.2-A x-ray crystal structure of Citrine, showing that the methionine side chain fills what was once a large halide-binding cavity adjacent to the chromophore. Insertion of calmodulin within Citrine or fusion of cyan Fluorescent Protein, calmodulin, a calmodulin-binding peptide and Citrine has generated improved calcium indicators. These chimeras can be targeted to multiple cellular locations and have permitted the first single-cell imaging of free [Ca 21 ] in the Golgi. Citrine is superior to all previous YFPs except when pH or halide sensitivity is desired and is particularly advantageous within genetically encoded Fluorescent indicators of physiological signals.
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circular permutation and receptor insertion within green Fluorescent Proteins
Proceedings of the National Academy of Sciences of the United States of America, 1999Co-Authors: Geoffrey S. Baird, David A Zacharias, Roger Y. TsienAbstract:Many areas of biology and biotechnology have been revolutionized by the ability to label Proteins genetically by fusion to the Aequorea green Fluorescent Protein (GFP). In previous fusions, the GFP has been treated as an indivisible entity, usually appended to the amino or carboxyl terminus of the host Protein, occasionally inserted within the host sequence. The tightly interwoven, three-dimensional structure and intricate posttranslational self-modification required for chromophore formation would suggest that major rearrangements or insertions within GFP would prevent fluorescence. However, we now show that several rearrangements of GFPs, in which the amino and carboxyl portions are interchanged and rejoined with a short spacer connecting the original termini, still become Fluorescent. These circular permutations have altered pKa values and orientations of the chromophore with respect to a fusion partner. Furthermore, certain locations within GFP tolerate insertion of entire Proteins, and conformational changes in the insert can have profound effects on the fluorescence. For example, insertions of calmodulin or a zinc finger domain in place of Tyr-145 of a Yellow mutant (enhanced Yellow Fluorescent Protein) of GFP result in indicator Proteins whose fluorescence can be enhanced severalfold upon metal binding. The calmodulin graft into enhanced Yellow Fluorescent Protein can monitor cytosolic Ca2+ in single mammalian cells. The tolerance of GFPs for circular permutations and insertions shows the folding process is surprisingly robust and offers a new strategy for creating genetically encodable, physiological indicators.
