Directly Measure CAR T Cell-mediated Cytotoxicity using Direct Cell Counting in 96-well Plates
- Scan the same plate over multiple time points to directly monitor the killing of target cells.
- Perform whole-well imaging and analysis of an entire 96-well plate in under 15 minutes
- No need trypsinize the cells and then perform lengthy flow cytometry protocols. The Celigo directly counts and reports every remaining target cell in each well
Introduction to CAR T Cell-Mediated Cytotoxicity
The Chimeric antigen receptor (CAR-T) technology focuses on generating a genetically modified t-cells that can directly bind to, and attack cells of interest. The CAR-T cell is comprised of an antigen-specific binding domain that is fused to the genetically engineered activating motif. Once expressed on the cell surface the CAR-T-cell can then target and attack the specific targeted cells.
- Target cells (either stably expressing GFP or labeled with a fluorescent dye) are seeded in the wells of microplates
- CAR-T cells were added to the wells at different Effector:Target (E:T) cell ratios
- Whole well images are acquired for each channel and the images are automatically analyzed to report the number of remaining GFP positive target cells