Cell Biology

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    Protocols in Current Issue
    Analysis of Caenorhabditis elegans Sperm Number, Size, Activation, and Mitochondrial Content
    Authors:  Amy M. Hammerquist, Chia-An Yen and Sean P. Curran, date: 06/05/2021, view: 236, Q&A: 0
    [Abstract]

    Infertility is a widespread and often unexplained issue. Studying reproduction using C. elegans males offers insight into the influence of individual factors on male fertility in humans. We have created a collection of protocols to assess several aspects of C. elegans sperm quality, including number, size, rate of activation, and mitochondrial

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    In vivo Fluorescence Imaging of Extracellular ATP in the Mouse Cerebral Cortex with a Hybrid-type Optical Sensor
    [Abstract]

    Adenosine 5’-triphosphate (ATP) works as an extracellular signaling molecule for cells in the brain, such as neurons and glia. Cellular communication via release of ATP is involved in a range of processes required for normal brain functions, and aberrant communication is associated with brain disorders. To investigate the mechanisms

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    Imaging and Fluorescence Quantification in Caenorhabditis elegans with Flow Vermimetry and Automated Microscopy
    Authors:  Elissa Tjahjono, Alexey V. Revtovich and Natalia V. Kirienko, date: 05/20/2021, view: 670, Q&A: 0
    [Abstract]

    Gene activation and cellular biomarkers are commonly monitored using fluorescent signals from transgenic reporters or dyes. These quantifiable markers are critical for biological research and serve as an incredibly powerful tool, even more so when combined with high-throughput screening. Caenorhabditis elegans is a particularly useful model in

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    Live Cell FRET Analysis of the Conformational Changes of Human P-glycoprotein
    Authors:  Ryota Futamata, Noriyuki Kioka and Kazumitsu Ueda, date: 02/20/2021, view: 1377, Q&A: 0
    [Abstract]

    The molecular mechanisms of P-glycoprotein (P-gp; also known as MDR1 or ABCB1) have been mainly investigated using artificial membranes such as lipid-detergent mixed micelles, artificial lipid bilayers, and membrane vesicles derived from cultured cells. Although these in vitro experiments help illustrate details about the molecular mechanisms of

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    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: 1188, 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: 1151, 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: 2169, 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: 1677, 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: 1606, 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, ...



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