Flash Sniper: Automated Detection and Analysis of Mitochondrial Superoxide Flash

摘要: Mitochondrial superoxide flash is a newly discovered physiological phenomenon reflecting elemental and bursting production in mitochondria of diverse cells culture vivo (Wang et al, Cell, 132, 279).The discovery opens unique window to glean into the mitochondrial ROS signaling its coupling with energy metabolism,cell fate regulation, oxidative stress-related diseases. Because low frequency long duration (∼20s) flash, time-lapse confocal imaging (xyt) was employed for data acquisition, resulting huge sets. For objective, reproducible efficient identification measurement, here we develop, validate implement an automated detection algorithm as well software, Flash Sniper, which fulfills three basic functions -- interactive priming, identification, parameter measurement. The priming module consists correction photobleach photoconversion using nonlinear fitting, cell motion compensation based on hierarchical block matching, spatiotemporal filtering. Two xy maps are generated represent maxima fluorescent intensity change corresponding time, respectively. events identified via thresholding segmentation these maps. Validation performed SNR-controlled synthetic sets produced by embedding typical real images. With aid examined ∼1600 flashes cardiac myocytes revealed distinct substructures their rising kinetics. In sum, analytical tool developed this study will facilitate studies mechanisms application health