Cell Biology

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    Protocols in Current Issue
    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: 593, 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: 484, 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 ...
    Flow Cytometry Analysis and Fluorescence-activated Cell Sorting of Myeloid Cells from Lung and Bronchoalveolar Lavage Samples from Mycobacterium tuberculosis-infected Mice
    Authors:  Alissa C. Rothchild, Dat Mai, Alan Aderem and Alan H. Diercks, date: 05/20/2020, view: 845, Q&A: 0
    [Abstract] Mycobacterium tuberculosis (Mtb) is transmitted by aerosol and can cause serious bacterial infection in the lung that can be fatal if left untreated. Mtb is now the leading cause of death worldwide by an infectious agent. Characterizing the early events of in vivo infection following aerosol challenge is critical for ...
    Evaluation of B Cell Proliferation in vivo by EdU Incorporation Assay
    Authors:  Adi Biram and Ziv Shulman, date: 05/05/2020, view: 744, Q&A: 0
    [Abstract] Generation of antibodies is crucial for establishing enduring protection from invading pathogens, as well as for maintaining homeostasis with commensal bacteria at mucosal surfaces. Chronic exposure to microbiota- and dietary- derived antigens results in continuous production of antibody producing cells within the Peyer’s patch germinal center ...
    Assessing in vitro and in vivo Trogocytosis By Murine CD4+ T cells
    Authors:  Jim Reed and Scott A. Wetzel, date: 05/05/2020, view: 706, Q&A: 0
    [Abstract] Recognition of antigens by lymphocytes (B, T, and NK) on the surface of an antigen-presenting cell (APC) leads to lymphocyte activation and the formation of an immunological synapse between the lymphocyte and the APC. At the immunological synapse APC membrane and associated membrane proteins can be transferred to the lymphocyte in a process called ...
    High-throughput Flow Cytometry Assay to Investigate TDP43 Splicing Function
    Authors:  H. Broder Schmidt and Rajat Rohatgi, date: 04/20/2020, view: 885, Q&A: 0
    [Abstract] Mutations in RNA-binding proteins (RBPs) such as TDP43 are associated with transcriptome-wide splicing defects and cause severe neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The impact of RBP mutations on splicing function is routinely studied using PCR-based bulk measurements. ...
    Dual Fluorescence Cytometry Assay to Assess Cellular Protein Levels
    Authors:  Ashley M Ngo, Ruofan Wang and Andreas S Puschnik, date: 04/20/2020, view: 930, Q&A: 0
    [Abstract] Expression levels of cellular proteins can be affected by various perturbations, such as genetic knockout of interactors, drug treatments or cell stress. To specifically measure the effects on protein levels post-synthesis under different experimental conditions, it is important to compensate for transcriptional and other upstream changes. Here, ...
    Flow Cytometry Measurement of Glucocerebrosidase Activity in Human Monocytes
    Authors:  Laura P. Hughes, Glenda M. Halliday and Nicolas Dzamko, date: 04/05/2020, view: 610, Q&A: 0
    [Abstract] Glucocerebrosidase (GCase) is an important enzyme for the metabolism of glycolipids. GCase enzyme deficiency is implicated in human disease and the efficient measurement of GCase activity is important for evaluating the efficacy of therapeutics targeting this enzyme. Existing approaches to measure GCase activity include whole blood mass ...
    Analyzing the Quenchable Iron Pool in Murine Macrophages by Flow Cytometry
    Authors:  Michael Riedelberger and Karl Kuchler, date: 03/20/2020, view: 631, Q&A: 0
    [Abstract] Tissue-resident macrophages are pivotal for a tightly-regulated iron metabolism at a cellular and systemic level, since subtle iron alterations increase the susceptibility for microbial infections or drive multiple diseases. However, research on cellular iron homeostasis in macrophages remains challenging due to the limited amount of available ...
    Growth Recovery Assay and FACS-based Population Sorting Following Territorial Exclusion in Proteus mirabilis
    Authors:  Murray J. Tipping and Karine A. Gibbs, date: 03/05/2020, view: 742, Q&A: 0
    [Abstract] Many bacteria take part in self recognition and kin discrimination behavior using contact-dependent effectors. Understanding the effects these effectors cause is important to explain bacterial community formation and population dynamics. Typically, kin discrimination effectors are toxins that kill target cells; their effect is therefore obvious ...



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