Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Proc Natl Acad Sci U S A. 2021 Jan 19;118(3). doi: 10.1073/pnas.2004176118.
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.
我们提出了一种基于压缩感知方案的活细胞高分辨率、高速荧光寿命成像显微镜(FLIM)。通过利用特定域中生物场景的可压缩性,我们在大视场范围内同时记录了脉冲激光激发后的时间分辨荧光衰减。该系统称为压缩 FLIM,可在单个相机曝光内获取全场荧光寿命图像,消除运动伪影并最小化光漂白和光毒性。成像速度仅受相机读出速度的限制,最高可达 100 Hz。我们展示了压缩 FLIM 在微观尺度上成像各种瞬态动力学的实用性。