Cell Biology

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    Protocols in Current Issue
    A Flow Cytometry-Based Method for Analyzing DNA End Resection in G0- and G1-Phase Mammalian Cells
    Authors:  Bo-Ruei Chen, Jessica K. Tyler and Barry P. Sleckman, date: 05/20/2022, view: 798, Q&A: 0
    [Abstract]

    DNA double strand breaks (DSBs) constantly arise in cells during normal cellular processes or upon exposure to genotoxic agents, and are repaired mostly by homologous recombination (HR) and non-homologous end joining (NHEJ). One key determinant of DNA DSB repair pathway choice is the processing of broken DNA ends to generate single strand DNA

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    Flow Cytometry-based Measurement of Reactive Oxygen Species in Cyanobacteria
    Authors:  Soumila Mondal and Shailendra P. Singh, date: 05/20/2022, view: 630, Q&A: 0
    [Abstract]

    Cyanobacteria are Gram-negative oxygen-producing photosynthetic bacteria that are useful in the pharmaceutical and biofuel industries. Monitoring of oxidative stress under fluctuating environmental conditions is important for determining the fitness, survival, and growth of cyanobacteria in the laboratory as well as in large scale cultivation

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    Small Molecule Screening of Primary Human Acute Myeloid Leukemia Using Co-culture and Multiplexed FACS Analysis
    Authors:  Aurelie Baudet, Simon Hultmark, Fredrik Ek and Mattias Magnusson, date: 03/20/2022, view: 560, Q&A: 0
    [Abstract]

    Ex vivo culture of primary acute myeloid leukemia (AML) cells is notoriously difficult due to spontaneous differentiation and cell death, which hinders mechanistic and translational studies. To overcome this bottleneck, we have implemented a co-culture system, where the OP9-M2 stromal cells support the growth, but most notably limit the

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    NBD-lipid Uptake Assay for Mammalian Cell Lines
    Authors:  Sara Abad Herrera, Marta Grifell-Junyent and Thomas Günther Pomorski, date: 02/20/2022, view: 1710, Q&A: 0
    [Abstract]

    All eukaryotic cells are equipped with transmembrane lipid transporters, which are key players in membrane lipid asymmetry, vesicular trafficking, and membrane fusion. The link between mutations in these transporters and disease in humans highlights their essential role in cell homeostasis. Yet, many key features of their activities, their

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    Flow Cytometry Analysis of Planarian Stem Cells Using DNA and Mitochondrial Dyes
    Authors:  Mohamed Mohamed Haroon, Praveen Kumar Vemula and Dasaradhi Palakodeti, date: 01/20/2022, view: 1111, Q&A: 0
    [Abstract]

    Planarians are free-living flatworms that emerged as a crucial model system to understand regeneration and stem cell biology. The ability to purify neoblasts, the adult stem cell population of planaria, through fluorescence-activated cell sorting (FACS) has tremendously increased our understanding of pluripotency, specialization, and

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    Evaluating Human Natural Killer Cells Antibody-dependent Cellular Cytotoxicity (ADCC) Using Plate-bound Anti-CD16 Antibodies
    Author:  Weiru Liu, date: 01/05/2022, view: 1352, Q&A: 0
    [Abstract]

    Natural killer (NK) cells are large granular lymphocytes that keep in check the health of neighboring cells through a large array of intrinsically expressed germline-coded receptors. Most importantly, CD16 is a low affinity Fc receptor for IgG that mediates the antibody-dependent cellular cytotoxicity (ADCC) of NK cells, bridging the innate and

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    Suppression of Human Dendritic Cells by Regulatory T Cells
    Authors:  Qing Huang, Avery J. Lam, Dominic A. Boardman, Nicholas A. J. Dawson and Megan K. Levings, date: 11/05/2021, view: 1694, Q&A: 0
    [Abstract]

    Regulatory T cells (Tregs) suppress immune responses via a variety of mechanisms and can be used as a cellular therapy to induce tolerance. The function of Tregs is commonly assessed in vitro using assays that measure suppression of effector T cell proliferation and/or cytokine production. However, Tregs can also suppress the function of antigen

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    Plasmodium cynomolgi Berok Growth Inhibition Assay by Thiol-reactive Probe Based Flow Cytometric Measurement
    Authors:  Jessica Jie Ying Ong, Bruce Russell and Jin-Hee Han, date: 09/05/2021, view: 1592, Q&A: 0
    [Abstract]

    The relapsing malaria species, Plasmodium vivax, is the most widely distributed and difficult-to-treat cause of human malaria. The merozoites of P. vivax preferentially invade ephemeral human CD71+ reticulocytes (nascent reticulocytes), thereby limiting the development of a robust continuous culture in vitro. Fortunately, P. vivax’s sister

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    One-step White Blood Cell Extracellular Staining Method for Flow Cytometry
    [Abstract]

    Flow cytometry is a powerful analytical technique that is increasingly used in scientific investigations and healthcare; however, it requires time-consuming, multi-step sample procedures, which limits its use to specialized laboratories. In this study, we propose a new universal one-step method in which white blood cell staining and red blood cell

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    Isolation of Microglia and Analysis of Protein Expression by Flow Cytometry: Avoiding the Pitfall of Microglia Background Autofluorescence
    Authors:  Jeremy C. Burns, Richard M. Ransohoff and Michaël Mingueneau, date: 07/20/2021, view: 3035, Q&A: 0
    [Abstract]

    Microglia are a unique type of tissue-resident innate immune cell found within the brain, spinal cord, and retina. In the healthy nervous system, their main functions are to defend the tissue against infectious microbes, support neuronal networks through synapse remodeling, and clear extracellular debris and dying cells through phagocytosis. Many

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