Cell Biology

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    Protocols in Current Issue
    Fibroblast Gap-closure Assay-Microscopy-based in vitro Assay Measuring the Migration of Murine Fibroblasts
    Authors:  Agnieszka P. Looney and Mallar Bhattacharya, date: 08/20/2019, view: 117, Q&A: 0
    [Abstract] Pulmonary fibrosis is characterized by pathological scaring of the lung. Similar to other fibrotic diseases, scar formation is driven by excessive extracellular matrix deposition by activated, proliferative, and migratory fibroblasts.

    Currently, the two most widely used chemotaxis and cell migration assays are the scratch assay and the ...
    Ex vivo Drosophila Wing Imaginal Disc Culture and Furin Inhibitor Assay
    Authors:  Alex Sohr, Lijuan Du and Sougata Roy, date: 08/20/2019, view: 52, Q&A: 0
    [Abstract] Furin is an evolutionarily conserved proprotein convertase (PC) family enzyme with a broad range of substrates that are essential for developmental, homeostatic, and disease pathways. Classical genetic approaches and in vitro biochemical or cell biological assays identified that precursor forms of most growth factor family proteins are ...
    A Novel Technique for Imaging and Analysis of Hair Cells in the Organ of Corti Using Modified Sca/eS and Machine Learning
    [Abstract] Here, we describe a sorbitol-based optical clearing method, called modified Sca/eS that can be used to image all hair cells (HCs) in the mouse cochlea. This modification of Sca/eS is defined by three steps: decalcification, de-lipidation, and refractive index matching, which can all be completed within 72 h. Furthermore, we established automated ...
    Cell-based Assay for Recruitment of DDR1 to Collagen-coated Beads
    Authors:  Victoria Juskaite and Birgit Leitinger, date: 08/20/2019, view: 64, Q&A: 0
    [Abstract] The discoidin domain receptors, DDR1 and DDR2, are key signaling receptors for the extracellular matrix protein collagen. The interactions of cells with collagen are difficult to study because of the difficulty to obtain native collagen fibers for in vitro studies. Thus, in vitro studies often use acid-soluble collagens in the ...
    Visualizing Hypoxia in a Murine Model of Candida albicans Infection Using in vivo Biofluorencence
    Authors:  José Pedro Lopes and Constantin F. Urban, date: 08/05/2019, view: 298, Q&A: 0
    [Abstract] Candida albicans is a leading human fungal pathogen that uses several metabolic adaptations to escape immune cells and causes systemic disease. Here, we describe a protocol for measuring one of these adaptations, the ability to thrive in hypoxic niches. Hypoxia was generated after successful subdermal infection with C. albicans ...
    Image-based Quantification of Direct Cell-to-cell Transmission of Bovine Viral Diarrhea Virus
    Authors:  Fernando Merwaiss and Diego E. Alvarez, date: 08/05/2019, view: 194, Q&A: 0
    [Abstract] Different viruses rely on direct cell-to-cell transmission to propagate infection within the infected host. Measuring this mode of transmission in cultured cells is often complicated by the contribution of cell free viruses to spread, and the difficulty to distinguish between primary infected cells that produce the virus and neighboring cells that ...
    QUEEN-based Spatiotemporal ATP Imaging in Budding and Fission Yeast
    Author:  Masak Takaine, date: 08/05/2019, view: 260, Q&A: 0
    [Abstract] Yeasts have provided an exceptional model for studying metabolism and bioenergetics in eukaryotic cells. Among numerous metabolites, adenosine triphosphate (ATP) is a major metabolite that is essential for all living organisms. Therefore, a clearer understanding of ATP dynamics in living yeast cells is important for deciphering cellular energy ...
    Lipid-exchange Rate Assay for Lipid Droplet Fusion in Live Cells
    Authors:  Jia Wang, Boon Tin Chua, Peng Li and Feng-Jung Chen, date: 07/20/2019, view: 484, Q&A: 0
    [Abstract] Lipid droplets (LDs) are central organelles in maintaining lipid homeostasis. Defective LD growth often results in the development of metabolic disorders. LD fusion and growth mediated by cell death–inducing DNA fragmentation factor alpha (DFFA)-like effector (CIDE) family proteins are crucial for various biological processes including unilocular ...
    Electron Microscopy Sample Preparation Protocol Enabling Nano-to-mesoscopic Mapping of Cellular Connectomes and Their Habitats in Human Tissues and Organs
    [Abstract] Multibeam scanning electron microscopy (multiSEM) provides a technical platform for seamless nano-to-mesoscale mapping of cells in human tissues and organs, which is a major new initiative of the U.S. National Institutes of Health. Such cross-length-scale imaging is expected to provide unprecedented understanding of relationships between cellular ...
    Live-cell Migration Assays to Study Motility of Neural and Glial (Oligodendrocyte) Progenitor Cells
    Authors:  Chu-Yen Chen, Fu-Sheng Chou and Pei-Shan Wang, date: 06/20/2019, view: 566, Q&A: 0
    [Abstract] Cell motility has been extensively studied in in vitro models using fibroblasts and keratocytes, but the cell type-specific mechanisms underlying migration of lineage- or disease-specific cells, such as neural and glial progenitor cells, remain an active field for investigation. The migrating neural and glial progenitor cells contribute ...



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