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    pH-Rodo pathogens to monitor phagocytosis in pMacs

    Rebecca Wallings

    Published: 2023-12-15 DOI: 10.17504/protocols.io.q26g7pn18gwz/v1
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    Abstract

    pH-Rodo pathogens to monitor phagocytosis in pMacs

    Steps

    1.

    Once pMacs have adhered to plate, resuspend pHrodoTM Green E. coli BioParticlesTM in InvitrogenTM Live Cell Imaging Solution to a final concentration of 1mg/mL and sonicate in bath sonicator for 10 minutes.

    Add 100 μL of E. coli BioParticles solution to each well (96-well plate).

    Place cells in a live imaging chamber (37°C, 5% carbon dioxide, and 95% humidity) of an EVOSTM M7000 (Invitrogen) at 20 x magnification capture 4 images per well every 20 minutes for 5 hours.

    Use brightfield images for automated-focus throughout the time course and use for normalization of GFP E. coli images.

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