Dec 15, 2023
DQ-BSA and BMV109 microscopy
- 1University of Florida
Protocol Citation: Rebecca Wallings 2023. DQ-BSA and BMV109 microscopy. protocols.io https://dx.doi.org/10.17504/protocols.io.261gedk5ov47/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: 92330
Funders Acknowledgement:
ASAP
Grant ID: ASAP-020527
Abstract
DQ-BSA and BMV109 microscopy for peritoneal macrophages
BMV109 Pan Cathepsin probe (Vergent Bioscience) and DQ red BSA (Invitrogen) were added to each well at a final concentration of 1µM and 10µG/mL, respectively, and cells were incubated at
37°C for 1-hour.
Cells were washed 3 x DPBS +/+ and fixed for 10 minutes at room temperature in Invitrogen‱ eBioscience‱ Intracellular Fixation buffer.
Cells were washed 3 x DPBS+/+ and incubated in 1 μg/ml DAPI (Life Technologies) for 10 minutes at room temperature in DPBS+/+.
Cells were imaged using an EVOS‱ M7000 (Invitrogen) at 20 x magnification. Image analysis was performed using Cellprofiler 4.2.5.