High-speed compressed-sensing fluorescence lifetime imaging microscopy of live cells

作者：Yayao Ma, Youngjae Lee, C. Best-Popescu, Liang Gao · 发表于：Proceedings of the National Academy of Sciences of the United States of America · 年份：2020 · DOI：10.1101/2020.07.16.205161 · 被引用次数：63 · 研究领域：Medicine、Materials Science、Biology

Significance This paper describes a high-speed fluorescence lifetime imaging method, compressed-sensing fluorescence lifetime imaging microscopy (compressed FLIM), which can produce high-resolution two-dimensional (2D) lifetime images at an unprecedented frame rate. Compared to other state-of-the-art FLIM imagers, compressed FLIM has a striking advantage in acquiring a widefield lifetime image within a single camera snapshot, thereby eliminating motion artifacts and enabling fast recording of biological events. We present high-resolution, high-speed fluorescence lifetime imaging microscopy (FLIM) of live cells based on a compressed sensing scheme. By leveraging the compressibility of biological scenes in a specific domain, we simultaneously record the time-lapse fluorescence decay upon pulsed laser excitation within a large field of view. The resultant system, referred to as compressed FLIM, can acquire a widefield fluorescence lifetime image within a single camera exposure, eliminating the motion artifact and minimizing the photobleaching and phototoxicity. The imaging speed, limited only by the readout speed of the camera, is up to 100 Hz. We demonstrated the utility of compressed FLIM in imaging various transient dynamics at the microscopic scale.