2025/01/10 by Zhihao Wei, Konglan Lin, Min Huang +2 · 1 voice
Immunology and Microbiology · Medicine · #CAR-T cell therapy research #Immune Cell Function and Interaction #T-cell and B-cell Immunology
paper · doi:10.1016/j.xpro.2024.103558
openalex publication_date 2025/01/10 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/03
Standard flow cytometry-based assays can determine the cytotoxicity of immune effector cells, but it is challenging to monitor the dynamic processes of cytotoxicity. Here, we present a protocol for continuous observation of natural killer (NK) cell-mediated cytotoxicity with microwell arrays using an automated microscope. We describe steps for isolating and labeling primary NK cells, loading cells onto microwell arrays, monitoring target wells, and image analysis. This protocol facilitates observation of the dynamics of immune-target cell interactions at the single-cell level. For complete details on the use and execution of this protocol, please refer to Li et al. 1 • Continuous observation of NK cell-mediated cytotoxicity at single-cell level • Real-time monitoring of NK-tumor cell interaction using an automated microscope • Quantification of NK cells that form immunological synapse with tumor cells Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Standard flow cytometry-based assays can determine the cytotoxicity of immune effector cells, but it is challenging to monitor the dynamic processes of cytotoxicity. Here, we present a protocol for continuous observation of natural killer (NK) cell-mediated cytotoxicity with microwell arrays using an automated microscope. We describe steps for isolating and labeling primary NK cells, loading cells onto microwell arrays, monitoring target wells, and image analysis. This protocol facilitates observation of the dynamics of immune-target cell interactions at the single-cell level.