Biological Sciences

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    Protocols in Current Issue
    Whole-mount Immunohistochemistry to Visualize Mouse Embryonic Dermal Lymphatic Vasculature
    Authors:  Xin Geng and R. Sathish Srinivasan, date: 10/20/2021, view: 125, Q&A: 0
    [Abstract]

    Lymphatic vessels are abundant in the skin where they regulate interstitial fluid uptake and immune surveillance. Defects in dermal lymphatic vessels, such as fewer vessels and abnormal lymphatic vessel coverage with mural cells, are frequently associated with lymphedema and other lymphatic disorders. Whole-mount immunohistochemistry allows the

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    High-throughput Screening for Defense Priming-inducing Compounds in Parsley Cell Cultures
    Authors:  Kathrin Schmitz, Linda Werner and Uwe Conrath, date: 10/20/2021, view: 121, Q&A: 0
    [Abstract]

    Defense priming describes the enhanced potency of cells to activate defense responses. Priming accompanies local and systemic immune responses and can be triggered by microbial infection or upon treatment with certain chemicals. Thus, chemically activating defense priming is promising for biomedicine and agriculture. However, test systems for

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    Quantification of RuBisCO Expression and Photosynthetic Oxygen Evolution in Cyanobacteria
    Authors:  Mateusz Kędzior and Betul Kacar, date: 10/20/2021, view: 154, Q&A: 0
    [Abstract]

    Phototrophic microorganisms are frequently engineered to regulate the expression and the activity of targeted enzymes of interest for specific biotechnological and agricultural applications. This protocol describes a method to evaluate the expression of RuBisCO (ribulose 1,5-bisphosphate carboxylase/oxygenase) in the model cyanobacterium

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    Zebrafish Embryos as a Predictive Animal Model to Study Nanoparticle Behavior in vivo
    Authors:  Gabriela Arias-Alpizar, Jeroen Bussmann and Frederick Campbell, date: 10/05/2021, view: 726, Q&A: 0
    [Abstract]

    A failure to fully understand the complex in vivo behavior of systemically administered nanomedicines has stymied clinical translation. To bridge this knowledge gap, new in vivo tools are needed to rapidly and accurately assess the nearly infinite array of possible nanoparticle designs. Zebrafish embryos are small, transparent, and easily

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    Inter-species Transplantation of Blastocysts between Medaka and Zebrafish
    Authors:  Jana F. Fuhrmann, Jasmin Onistschenko and Lázaro Centanin, date: 09/20/2021, view: 615, Q&A: 0
    [Abstract]

    Transplantation of blastocysts from a donor to a host blastula constitutes a powerful experimental tool to tackle major developmental biology questions. The technique is widely implemented in diverse biological models including teleost fish, where it is typically used for intra-species blastula transplantations – i.e., labeled blastocysts

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    Intrathoracic Inoculation of Zika Virus in Aedes aegypti
    Authors:  Irma Sanchez-Vargas, Adeline E. Williams, Alexander W.E. Franz and Ken E. Olson, date: 09/20/2021, view: 348, Q&A: 0
    [Abstract]

    Aedes aegypti mosquitoes are the main vectors of many medically relevant arthropod-borne (arbo) viruses, including Zika (ZIKV), dengue (DENV), and yellow fever (YFV). Vector competence studies with Ae. aegypti often involve challenging mosquitoes with an artificial bloodmeal containing virus and later quantifying viral titer or infectious

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    A Co-purification Method for Efficient Production and Src Kinase-mediated Phosphorylation of Aplysia Cortactin
    Authors:  Sherlene L. Brown, Yuan Ren, Daniel M. Suter and Seema Mattoo, date: 09/20/2021, view: 452, Q&A: 0
    [Abstract]

    Cortactin is an actin-binding protein that regulates processes like cell migration, endocytosis, and tumor cell metastasis. Although cortactin is associated with actin-cytoskeletal dynamics in non-neuronal cells and cell-free systems, the exact mechanisms underlying its fundamental roles in neuronal growth cones are not fully explored. Recent

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    In vitro Cleavage and Electrophoretic Mobility Shift Assays for Very Fast CRISPR
    Authors:  Roger S. Zou, Yang Liu and Taekjip Ha, date: 09/05/2021, view: 925, Q&A: 0
    [Abstract]

    CRISPR-Cas9 has transformed biomedical research and medicine through convenient and targeted manipulation of DNA. Time- and spatially-resolved control over Cas9 activity through the recently developed very fast CRISPR (vfCRISPR) system have facilitated comprehensive studies of DNA damage and repair. Understanding the fundamental principles of Cas9

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    Using Atomic Force Microscopy to Study the Real Time Dynamics of DNA Unwinding by Mitochondrial Twinkle Helicase
    Authors:  Parminder Kaur, Hai Pan, Matthew J. Longley, William C. Copeland and Hong Wang, date: 09/05/2021, view: 932, Q&A: 0
    [Abstract]

    Understanding the structure and dynamics of DNA-protein interactions during DNA replication is crucial for elucidating the origins of disorders arising from its dysfunction. In this study, we employed Atomic Force Microscopy as a single-molecule imaging tool to examine the mitochondrial DNA helicase Twinkle and its interactions with DNA. We used

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    Isolation and Characterization of Membrane Vesicles from Lactobacillus Species
    Authors:  Julie C. Caruana, Scott N. Dean and Scott A. Walper, date: 09/05/2021, view: 974, Q&A: 0
    [Abstract]

    Throughout their life cycle, bacteria shed portions of their outermost membrane comprised of proteins, lipids, and a diversity of other biomolecules. These biological nanoparticles have been shown to have a range of highly diverse biological activities, including pathogenesis, community regulation, and cellular defense (among others). In recent

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