(A) Near-atomic resolution studies of in vitro monolayer systems. After preparing the monolayer systems, which may contain signaling components carrying functional mutations (ON, wild-type, OFF), a standard procedure is used to prepare samples for the microscope. (B) A proposed workflow for time-resolved studies of lysed cells and minicells. After preparing the biological samples, a FRET-based kinase assay is used to determine functionally interesting time points based on chemosensory array kinetics (e.g. in response to treatment with a chemoeffector). Grid samples are then exposed to stimulus in a timed fashion (e.g. through the photolysis of a caged ligand) and vitrified at chosen time points to capture signaling states of interest. (C) Structure determination for both protocols follows a similar workflow based on a standard cryoET structure analysis pipeline, complemented by computational modeling techniques.