Towards fully automated Identification of Vesicle‐Membrane Fusion Events in TIRF Microscopy

Total Internal Reflection Fluorescence Microscopy (TIRFM) is imposing itself as the tool of choice for studying biological activity in close proximity to the plasma membrane. For example, the exquisite selectivity of TIRFM allows monitoring the diffusion of GFP‐phogrin vesicles and their recruitment to the plasma membrane in pancreatic β‐cells. We present a novel computer vision system for automatically identifying the elusive fusion events of GFP‐phogrin vesicles with the plasma membrane. Our method is based on robust object tracking and matched filtering. It should accelerate the quantification of TIRFM data and allow the extraction of more biological information from image data to support research in diabetes and obesity.