Cell Biology

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    Protocols in Current Issue
    Live-cell Imaging and Quantitative Analysis of Meiotic Divisions in Caenorhabditis elegans Males
    Authors:  Gunar Fabig, Falko Löffler, Christian Götze and Thomas Müller-Reichert, date: 10/20/2020, view: 363, Q&A: 0
    [Abstract] Live-imaging of meiotic cell division has been performed in extracted spermatocytes of a number of species using phase-contrast microscopy. For the nematode Caenorhabditis elegans, removal of spermatocytes from gonads has damaging effects, as most of the extracted spermatocytes show a high variability in the timing of meiotic divisions or ...
    Fluorescence Measurement and Calibration of Intracellular pH in Starfish Oocytes
    Authors:  Enako Hosoda and Kazuyoshi Chiba, date: 10/05/2020, view: 336, Q&A: 0
    [Abstract] Oocyte maturation is a process wherein an oocyte arrested at prophase of meiosis I resumes meiosis to become a fertilizable egg. In starfish ovaries, a hormone released from follicle cells activates the oocytes, resulting in an increase in their intracellular pH (pHi), which is required for spindle assembly. Herein, we describe a ...
    Site-specific Labeling of B Cell Receptor and Soluble Immunoglobulin
    Authors:  Yu Wang, Zhixun Shen, Zhengpeng Wan and Wanli Liu, date: 09/20/2020, view: 757, Q&A: 0
    [Abstract] B lymphocyte activation is regulated by its membrane-bound B cell receptors (BCRs) upon recognizing diverse antigens. It is hypothesized that antigen binding would trigger conformational changes within BCRs, followed by a series of downstream signaling activation. To measure the BCR conformational changes in live cells, a fluorescent site-specific ...
    Quantitative Kinetic Analyses of Histone Turnover Using Imaging and Flow Cytometry
    Authors:  Hanae Sato, Robert H. Singer and John M. Greally, date: 09/05/2020, view: 916, Q&A: 0
    [Abstract] Dynamic histone changes occur as a central part of chromatin regulation. Deposition of histone variants and post-translational modifications of histones are strongly associated with properties of chromatin status. Characterizing the kinetics of histone variants allows important insights into transcription regulation, chromatin maintenance and ...
    In vitro Cultivation and Visualization of Malaria Liver Stages in Primary Simian Hepatocytes
    Authors:  Devendra Kumar Gupta and Thierry Diagana, date: 08/20/2020, view: 805, Q&A: 0
    [Abstract] Human liver is the primary and obligatory site for malaria infection where sporozoites invade host hepatocytes. Malaria hepatic stages are asymptomatic and represent an attractive target for development of anti-malarial interventions and vaccines. However, owing to lack of robust and reproducible in vitro culture system, it is difficult ...
    Measuring Intracellular Vesicle Density and Dispersion Using Fluorescence Microscopy and ImageJ/FIJI
    [Abstract] Cell signalling, cell secretion, and plasma membrane repair are processes that critically rely on intracellular vesicles, important components of the endocytic and secretory pathways. More specifically, the strategic distribution of intracellular vesicles is important for diverse cellular processes. The method presented here is a simple, ...
    Preparation of Drosophila Polytene Chromosomes, Followed by Immunofluorescence Analysis of Chromatin Structure by Multi-fluorescence Correlations
    Authors:  Terra M. Kuhn, Shawn C. Little and Maya Capelson, date: 07/05/2020, view: 1306, Q&A: 0
    [Abstract] Drosophila larval salivary gland polytene chromosome squashes have been used for decades to analyze genome-wide protein-binding patterns, transcriptional activation processes, and changes in chromatin structure at specific genetic loci. There have been many evolutions of the squashing protocol over the years, with sub-optimal ...
    Single Cell Volume Measurement Utilizing the Fluorescence Exclusion Method (FXm)
    Authors:  Nash D. Rochman, Kai Yao, Nicolas A. Perez Gonzalez, Denis Wirtz and Sean X. Sun, date: 06/20/2020, view: 1187, Q&A: 0
    [Abstract] The measurement of single cell size remains an obstacle towards a deeper understanding of cell growth control, tissue homeostasis, organogenesis, and a wide range of pathologies. Recent advances have placed a spotlight on the importance of cell volume in the regulation of fundamental cell signaling pathways including those known to orchestrate ...
    Live Cell Measurement of the Intracellular pH of Yeast by Flow Cytometry Using a Genetically-Encoded Fluorescent Reporter
    Authors:  Catherine G. Triandafillou and D. Allan Drummond, date: 06/20/2020, view: 1100, Q&A: 0
    [Abstract] The intracellular pH of yeast is a tightly regulated physiological cue that changes in response to growth state and environmental conditions. Fluorescent reporters, which have altered fluorescence in response to local pH changes, can be used to measure intracellular pH. While microscopy is often used to make such measurements, it is relatively ...
    Live Mitochondrial or Cytosolic Calcium Imaging Using Genetically-encoded Cameleon Indicator in Mammalian Cells
    Authors:  Elisa Greotti and Tullio Pozzan, date: 02/05/2020, view: 2155, Q&A: 0
    [Abstract] Calcium (Ca2+) imaging aims at investigating the dynamic changes in live cells of its concentration ([Ca2+]) in different pathophysiological conditions. Ca2+ is an ubiquitous and versatile intracellular signal that modulates a large variety of cellular functions thanks to a cell type-specific toolkit and a complex ...



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