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    Protocols in Current Issue
    Brain-localized and Intravenous Microinjections in the Larval Zebrafish to Assess Innate Immune Response
    Authors:  Alison M. Rojas and Celia E. Shiau, date: 04/05/2021, view: 611, Q&A: 0
    [Abstract]

    Creating a robust and controlled infection model is imperative for studying the innate immune response. Leveraging the particular strengths of the zebrafish model system, such as optical transparency, ex utero development, and large clutch size, allows for the development of methods that yield consistent and reproducible results. We created a

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    Liposomal Clodronate-mediated Macrophage Depletion in the Zebrafish Model
    Authors:  Linlin Yang, Alison M. Rojas and Celia E. Shiau, date: 03/20/2021, view: 833, Q&A: 0
    [Abstract]

    The ability to conduct in vivo macrophage-specific depletion remains an effective means to uncover functions of macrophages in a wide range of physiological contexts. Compared to the murine model, zebrafish offer superior imaging capabilities due to their optical transparency starting from a single-cell stage to throughout larval development.

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    Automated Analysis of Cerebrospinal Fluid Flow and Motile Cilia Properties in The Central Canal of Zebrafish Embryos
    [Abstract]

    Circulation of cerebrospinal fluid (CSF) plays an important role during development. In zebrafish embryo, the flow of CSF has been found to be bidirectional in the central canal of the spinal cord. In order to compare conditions and genetic mutants across each other, we recently automated the quantification of the velocity profile of exogenous

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    Whole-mount Immunohistochemistry of Adult Zebrafish Retina for Advanced Imaging
    Authors:  Mikiko Nagashima and Peter F. Hitchcock, date: 12/20/2020, view: 1159, Q&A: 0
    [Abstract]

    Immunohistochemistry is a widely used technique to examine the expression and subcellular localization of proteins. This technique relies on the specificity of antibodies and requires adequate penetration of antibodies into tissues. The latter is especially challenging for thick specimens, such as embryos and other whole-mount preparations. Here

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    Use of Optogenetic Amyloid-β to Monitor Protein Aggregation in Drosophila melanogaster, Danio rerio and Caenorhabditis elegans
    Authors:  Prameet Kaur, Caroline Kibat, Emelyne Teo, Jan Gruber, Ajay Mathuru and Nicholas S. Tolwinski, date: 12/05/2020, view: 1229, Q&A: 0
    [Abstract]

    Alzheimer’s Disease (AD) has long been associated with accumulation of extracellular amyloid plaques (Aβ) originating from the Amyloid Precursor Protein. Plaques have, however, been discovered in healthy individuals and not all AD brains show plaques, suggesting that extracellular Aβ aggregates may play a smaller role than

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    Advances in Proximity Ligation in situ Hybridization (PLISH)
    Authors:  Monica Nagendran, Adam M Andruska, Pehr B Harbury and Tushar J Desai, date: 11/05/2020, view: 1637, Q&A: 0
    [Abstract] Understanding tissues in the context of development, maintenance and disease requires determining the molecular profiles of individual cells within their native in vivo spatial context. We developed a Proximity Ligation in situ Hybridization technology (PLISH) that enables quantitative measurement of single cell gene expression ...
    A Reproducible Protocol to Measure the Critical Swimming Speed of Adult Zebrafish
    Authors:  Yuma Wakamatsu, Makoto Kashima and Hiromi Hirata, date: 08/20/2020, view: 1747, Q&A: 1
    [Abstract] The quantitative measurement of water flow-induced swimming of fish species using a swimmill is a powerful method to evaluate motor ability of individual fish. Zebrafish is a commonly used vertebrate that enables the study of morphological, physiological and behavioral characteristics associated with genes. We here established a reproducible ...
    Paper Lateral Flow Biosensor for Nodavirus Reverse Transcribed RNA Detection
    Authors:  Dimitra K. Toubanaki and Evdokia Karagouni, date: 08/05/2020, view: 1113, Q&A: 0
    [Abstract] Paper nanobiosensors have been established as an excellent platform for analysis of veterinary and human pathogens causing various diseases. Especially, lateral flow assays or biosensors ideal for sensitive, rapid, robust and accurate analysis in laboratory setups and on-site analysis. Viral RNA detection is of great importance for public health ...
    Low-cost and Multiplexable Whole mRNA-Seq Library Preparation Method with Oligo-dT Magnetic Beads for Illumina Sequencing Platforms
    Authors:  Makoto Kashima, Ayumi Deguchi, Ayumi Tezuka and Atsushi J. Nagano, date: 06/20/2020, view: 2769, Q&A: 0
    [Abstract] RNA-Seq is a powerful method for transcriptome analysis used in varied field of biology. Although several commercial products and hand-made protocols enable us to prepare RNA-Seq library from total RNA, their cost are still expensive. Here, we established a low-cost and multiplexable whole mRNA-Seq library preparation method for illumine ...
    Quantitative Determination of Ca2+-binding to Ca2+-sensor Proteins by Isothermal Titration Calorimetry
    Authors:  Seher Abbas and Karl-Wilhelm Koch, date: 04/05/2020, view: 2086, Q&A: 0
    [Abstract] Diverse and complex molecular recognitions are central elements of signal transduction cascades. The strength and nature of these interaction modes can be determined by different experimental approaches. Among those, Isothermal titration calorimetry (ITC) offers certain advantages by providing binding constants and thermodynamic parameters from ...



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