Dec 15, 2023
pH-Rodo pathogens to monitor phagocytosis in pMacs
- 1University of Florida
Protocol Citation: Rebecca Wallings 2023. pH-Rodo pathogens to monitor phagocytosis in pMacs. protocols.io https://dx.doi.org/10.17504/protocols.io.q26g7pn18gwz/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 14, 2023
Last Modified: December 15, 2023
Protocol Integer ID: 92331
Funders Acknowledgement:
ASAP
Grant ID: ASAP-020527
Abstract
pH-Rodo pathogens to monitor phagocytosis in pMacs
Once pMacs have adhered to plate, resuspend pHrodo‱ Green E. coli BioParticles‱ in Invitrogen‱ 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‱ 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.