ABSTRACT Cell and gene therapies have revolutionized treatment options for patients with pace of discovery and implementation accelerating. For example, clinical success of CAR-T cells for hematologic malignancies has led to use in expanded patient populations and trials targeting for other cancer types. While there are many cell therapies in preclinical development and clinical trials confidence in their successes is undermined by the inherent complexity of cells that remains difficult to decipher using existing tools. What is needed is an integrated analytical platform providing a matrix of readouts that reveal an in-depth real-time understanding of cell phenotype and metabolism. Such a platform will inform on the therapeutic potential before administration and by sampling patients will reveal aspects of cell behavior that correlate with efficacy. CellChorus is answering this need using our platform (“TIMING”) to undertake large scale single-cell analyses to track and characterize behavior of immune cells over time. TIMING uses sophisticated AI-based software, many years in development, to track and characterize behaviors of thousands of cells in parallel using time-lapse imaging. Multiple papers and repeat customers have helped validate our approach to commercialization. Based on users’ demand, we propose to advance TIMING by (i) developing and optimizing reagents and protocols to identify a suite of cell surface attributes, (ii) quantify proliferative capacity and (iii) determine metabolic activity before, during, and after cell-cell interactions. We will integrate these findings with our AI software to provide end users with a simple, unified assay system to run in their own laboratories on commercially available imaging instrumentation. The results of this project will be ready-to-use kits to reveal critical immune-biology of cells in critical and challenging environments. This is predicted to augment translational research and development leading to improvements in manufacturing and success in patients.