After infecting iPSC-cardiomyocytes with GCaMP6f for a calcium transience assay using a field stimulation pacer at 0.2 Hz, I notice the cells dying after about 6-12 hours. The auto microscope used has an incubator that is both CO2 and temperature controlled. Over time, the transients per minute significantly increase and the cells eventually die. Does anyone have any experience with this assay/ might have insight into why this is occurring and potential solutions? The pH of the media seems to remain the same and no visible contamination is observed.