PROJECT ABSTRACT Nearly two thirds of the 18 million new cancer patients each year fail treatment due to resistance that is not fully understood. Even in state-of-the-art clinical trials, oncologists struggle to select patients likely to respond to next-generation therapies. The outdated, imprecise pathology tools currently in clinical use (e.g. IHC) fail to reliably deliver critical information such as the extent of drug target expression and accurate assessment of cell states. Newer research tools including immunofluorescence analysis and mass cytometry also fail to bridge this gap due to complex workflows that limit adoption to expert users, incur high sample size requirements, and/or represent a prohibitive expense. There is a significant unmet clinical need for the development of an improved diagnostic tool capable of generating precise, accurate, single-cell information to enable improved diagnostic and treatment outcomes across tumor types. The goal of this project is to commercialize a platform for rapid molecular analysis for minute and fragile cell populations, in order to generate data on protein expression, cell state, and co-localization (including of immune cells) in single cells with the precision of flow cytometry, while preserving the sample. The successful completion of this project will result in a validated panel for profiling of immune cells with accompanying computational services, which can be used on standard immunofluorescent microscopes. A successful project of this service is expected to translate into the ability to profile many more biomarkers from a range of biopsy samples than is possible at present. Ultimately, the technology is well suited to translate to clinical settings, where it can be used to optimally select responders for precision cancer therapies.