Images were collected using the open-source Micromanager image acquisition software. Camera settings were standardized and optimized by cooling the chip to the lowest temperature (to minimize dark current) and maximizing the gain. Binning was used only when it was necessary to produce visible images. All of the background light sources in the room (windows, doors, and electronics) were covered with blackout material and images were collected at various exposure times (1 to 40 s). Long-term bioluminescence images were acquired using the multiacquisition feature of Micromanager and a perfusion system to deliver CTZ during the imaging period (up to 3 h). Imaging was done under controlled conditions that included using the same batch of transfected cells, CTZ concentration, media, exposure times, and imaging conditions to reduce variability between experiments. We did not attempt to normalize bioluminescence signals between different sets of experiments, although we employed the same camera settings (i.e., exposure time and binning) for a given combination of the microscope, camera, objective lens, and luciferase used. Thus, the bioluminescence signal was comparable within a given set of the experiments.