Mitochondrial apoptosis imaged by high-throughput confocal microscopy

Mitochondrial programmed cell death (i.e. intrinsic apoptosis) is regulated through the formation of pro-apoptotic protein complexes at the mitochondrial outer membrane (MOM), which permeabilize the MOM and commit the cell to die. This step is modulated by mitochondrial dynamics. To capture this, we treated cells with the mitochondrial dynamics inhibitor M1 before inducing apoptosis with actinomycin D (ActD), and imaged them by high-throughput spinning-disk confocal microscopy. In mouse embryonic fibroblasts (MEFs) we followed apoptosis by long-term live-cell imaging, and in human induced pluripotent stem cells (hiPSCs) we stained fixed cells for the pro-apoptotc protein BAX at mitochondria. Here, we provide a description of these two large-scale microscopy datasets. Dataset 1 contains 637,632 images of MEFs imaged as time series after labelling of mitochondrial morphology and integrity with the mitochondrial membrane potential-sensitive dye tetramethylrhodamine-ethylester (TMRE) and apoptosis with Annexin V-FITC. The dataset also contains a digital phase contrast channel which can be useful to track cells. Dataset 2 contains 9,504 images of hiPSCs with immunofluorescence labelling of BAX and COX IV. The datasets are made available through the BioImage Archive repository.