pH-Rodo pathogens to monitor phagocytosis in pMacs

Once pMacs have adhered to plate, resuspend pHrodo^TM Green E. coli BioParticles^TM in Invitrogen^TM 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 EVOS^TM 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.