Development of a Workflow for a Robust Cytometry of Reaction Rate Constant

Cytometry of Reaction Rate Constant (CRRC) is a method for studying cell population heterogeneity based on the activity of cellular processes. The original CRRC workflow did not account for cell motility, which led to inaccurate measurements in motile cells. Here, we report on the development of a new CRRC workflow that makes it applicable to motile cells. We confirmed the robustness of the new workflow to cell movement by performing a comparative CRRC workflow study of cross-membrane transport in motile cells. Using the new workflow, preliminary progress was made on the investigation of cytochrome p450 (CYP) activity. We validated the CRRC experimental procedure to conduct such study but found that the CRRC CYP assay had considerable variability. Nonetheless, the development of the new CRRC workflow is a step in the right direction with more work needing to be done to understand the variability found in the CRRC CYP assay.