Event-based vision sensor for fast and dense single-molecule localization microscopy

Single-molecule localization microscopy (SMLM) enables crucial insights into cellular structures and processes to be revealed at the single-molecule level. However, SMLM is often hampered by limited temporal resolution and the fixed frame rate of the acquisition. Here we present a new approach to SMLM data acquisition and processing based on an affordable event-based sensor. This type of sensor reacts to changes in light intensity, rather than integrating photons during the exposure time of each frame. Each pixel works independently and returns a signal only when an intensity change is detected. Compared with video acquisition using traditional electron-multiplying charge-coupled device or scientific complementary metal–oxide–semiconductor cameras, the event-based sensor provides higher temporal resolution and throughput on the positions of blinking molecules. We demonstrate event-based SMLM super-resolution imaging on biological samples with spatial resolution on a par with the performance of electron-multiplying charge-coupled device or scientific complementary metal–oxide–semiconductor cameras, while registering only the on and off switching of blinking molecules. We use event-based SMLM to perform very dense single-molecule imaging, where frame-based cameras experience major limitations. Event-based sensors enable super-resolution single-molecule localization microscopy with comparable quality and resolution to traditional scientific cameras, while also overcoming the limitations of high-density imaging.