Cell Biology

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    Protocols in Current Issue
    Production of Phenotypically Uniform Human Cerebral Organoids from Pluripotent Stem Cells
    Authors:  Adam Sivitilli, Parisa Ghiasi and Liliana Attisano, date: 04/20/2021, view: 110, Q&A: 0

    Recent advances in stem cell technology have allowed researchers to generate 3D cerebral organoids (COs) from human pluripotent stem cells (hPSCs). Indeed, COs have provided an unprecedented opportunity to model the developing human brain in a 3D

    ...
    AIMTOR, a BRET Biosensor for Live Recording of mTOR Activity in Cell Populations and Single Cells
    Authors:  Nathalie Bouquier, Enora Moutin, Julie Perroy and Vincent Ollendorff, date: 04/20/2021, view: 71, Q&A: 0

    Mammalian target of rapamycin (mTOR) controls many crucial cellular functions, including protein synthesis, cell size, energy metabolism, lysosome and mitochondria biogenesis, and autophagy. Consequently, deregulation of mTOR signaling plays a role

    ...
    Measuring Cytosolic Translocation of Mycobacterium marinum in RAW264.7 Macrophages with a CCF4-AM FRET Assay
    Authors:  Javier Aguilera and Jianjun Sun, date: 04/20/2021, view: 79, Q&A: 0
    The CCF4-AM Förster resonance energy transfer (FRET) assay is a sensitive approach to measure bacterial cytosolic translocation in live cells. The FRET pair hydroxycoumarin (donor) and fluorescein (acceptor) are linked by a CCF4-AM β-lactam ring, ...
    Single Cell Analysis and Sorting of Aspergillus fumigatus by Flow Cytometry
    Authors:  Gareth Howell and Robert-Jan Bleichrodt, date: 04/20/2021, view: 76, Q&A: 0

    Experimental results in fungal biology research are usually obtained as average measurements across whole populations of cells, whilst ignoring what is happening at the single cell level. Microscopy has allowed us to study single-cell behavior, but

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    A Robust Mammary Organoid System to Model Lactation and Involution-like Processes
    Authors:  Elsa Charifou, Jakub Sumbal, Zuzana Koledova, Han Li and Aurélie Chiche, date: 04/20/2021, view: 201, Q&A: 0

    The mammary gland is a highly dynamic tissue that changes throughout reproductive life, including growth during puberty and repetitive cycles of pregnancy and involution. Mammary gland tumors represent the most common cancer diagnosed in women

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    A Robust Mammary Organoid System to Model Lactation and Involution-like Processes
    Authors:  Elsa Charifou, Jakub Sumbal, Zuzana Koledova, Han Li and Aurélie Chiche, date: 04/20/2021, view: 201, Q&A: 0
    [Abstract]

    The mammary gland is a highly dynamic tissue that changes throughout reproductive life, including growth during puberty and repetitive cycles of pregnancy and involution. Mammary gland tumors represent the most common cancer diagnosed in women worldwide. Studying the regulatory mechanisms of mammary gland development is essential for understanding

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    Single Cell Analysis and Sorting of Aspergillus fumigatus by Flow Cytometry
    Authors:  Gareth Howell and Robert-Jan Bleichrodt, date: 04/20/2021, view: 76, Q&A: 0
    [Abstract]

    Experimental results in fungal biology research are usually obtained as average measurements across whole populations of cells, whilst ignoring what is happening at the single cell level. Microscopy has allowed us to study single-cell behavior, but it has low throughput and cannot be used to select individual cells for downstream experiments. Here

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    Measuring Cytosolic Translocation of Mycobacterium marinum in RAW264.7 Macrophages with a CCF4-AM FRET Assay
    Authors:  Javier Aguilera and Jianjun Sun, date: 04/20/2021, view: 79, Q&A: 0
    [Abstract] The CCF4-AM Förster resonance energy transfer (FRET) assay is a sensitive approach to measure bacterial cytosolic translocation in live cells. The FRET pair hydroxycoumarin (donor) and fluorescein (acceptor) are linked by a CCF4-AM β-lactam ring, the substrate of β-lactamase. The exogenously added, neutral charged-FRET reagent can diffuse across ...
    AIMTOR, a BRET Biosensor for Live Recording of mTOR Activity in Cell Populations and Single Cells
    Authors:  Nathalie Bouquier, Enora Moutin, Julie Perroy and Vincent Ollendorff, date: 04/20/2021, view: 71, Q&A: 0
    [Abstract]

    Mammalian target of rapamycin (mTOR) controls many crucial cellular functions, including protein synthesis, cell size, energy metabolism, lysosome and mitochondria biogenesis, and autophagy. Consequently, deregulation of mTOR signaling plays a role in numerous pathological conditions such as cancer, metabolic disorders and neurological diseases.

    ...
    Production of Phenotypically Uniform Human Cerebral Organoids from Pluripotent Stem Cells
    Authors:  Adam Sivitilli, Parisa Ghiasi and Liliana Attisano, date: 04/20/2021, view: 110, Q&A: 0
    [Abstract]

    Recent advances in stem cell technology have allowed researchers to generate 3D cerebral organoids (COs) from human pluripotent stem cells (hPSCs). Indeed, COs have provided an unprecedented opportunity to model the developing human brain in a 3D context, and in turn, are suitable for addressing complex neurological questions by leveraging

    ...
    A Workflow for High-pressure Freezing and Freeze Substitution of the Caenorhabditis elegans Embryo for Ultrastructural Analysis by Conventional and Volume Electron Microscopy
    Authors:  Mohammad M. Rahman, Irene Y. Chang, Orna Cohen-Fix and Kedar Narayan, date: 04/05/2021, view: 250, Q&A: 0
    [Abstract]

    The free-living nematode Caenorhabditis elegans is a popular model system for studying developmental biology. Here we describe a detailed protocol to high-pressure freeze the C. elegans embryo (either ex vivo after dissection, or within the intact worm) followed by quick freeze substitution. Processed samples are suitable for ultrastructural

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    Ex vivo Assessment of Mitochondrial Function in Human Peripheral Blood Mononuclear Cells Using XF Analyzer
    Authors:  Alica Schöller-Mann, Katja Matt, Barbara Hochecker and Jörg Bergemann, date: 04/05/2021, view: 182, Q&A: 0
    [Abstract]

    Cellular health and function, as we know today, depend on a large extent on mitochondrial function. The essential function of mitochondria is the energy production, more precisely ATP production, via oxidative phosphorylation. Mitochondrial energy production parameters therefore represent important biomarkers. Studies on human cells have mainly

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    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: 372, 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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    Measurements of Root Colonized Bacteria Species
    Authors:  Vílchez Juan Ignacio, Yu Yang, Dian Yi and Huiming Zhang, date: 04/05/2021, view: 299, Q&A: 0
    [Abstract]

    Root-associated bacteria are able to influence plant fitness and vigor. A key step in understanding the belowground plant-bacteria interactions is to quantify root colonization by the bacteria of interest. Probably, genetic engineering with fluorescence markers is the most powerful way to monitor bacterial strains in plant. However, this could

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    Investigate Synaptic Vesicles Mobility in Neuronal Culture Axons by FRAP Imaging
    Authors:  Xiao Min Zhang, Fabrice P. Cordelieres and Etienne Herzog, date: 03/20/2021, view: 615, Q&A: 0
    [Abstract]

    Synaptic vesicles (SVs) are clustered in the presynaptic terminals and consistently trafficking along axons. Based on their release features, SVs are classified into different “pools”. Imaging of SVs that are traveling among multiple presynaptic terminals has helped define a new pool named “SV super-pool”. Here we describe

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