The footprinting method Fast Photochemical Oxidation of proteins (FPOP) has been extended to live cells in a method entitled in-cell FPOP (IC-FPOP). Owing to microsecond time scale of labeling, IC-FPOP is suited to study fast biological processes in the complex cellular environment. While a single cell flow system laid the foundation for the success of IC-FPOP, its temporal limitation motivated the design of a higher throughput platform, Platform Incubator with Movable XY stage (PIXY). Advancements have been made to automate the PIXY system. Currently, validation studies strive to address the utility of automation for IC-FPOP. This current study validates PIXY’s ability to accommodate automated time points and subsequent changes over time targeting the endoplasmic reticulum stress pathway.