National Research Council of Canada, Steacie Institute for Molecular Sciences, Ottawa, Ontario, K1A 0R6 Canada.
J Biomed Opt. 2011 Feb;16(2):021103. doi: 10.1117/1.3490641.
We demonstrate the simultaneous collection and separation of femtosecond-laser-based forward-collected coherent anti-Stokes Raman scattering (F-CARS) and two-photon-excitation-induced fluorescence lifetime images (FLIM) using time-correlated single photon counting (TCSPC). We achieve this in a nondescanned geometry using a single multimode fiber without significant loss of light, field of view, and most importantly, TCSPC timing fidelity. In addition to showing the ability to separate CARS images from FLIM images using time gating, we also demonstrate composite multimodal epicollected FLIM imaging with fiber-collected F-CARS imaging in live cells.
我们展示了使用时间相关单光子计数 (TCSPC) 同时采集和分离飞秒激光基正向收集相干反斯托克斯拉曼散射 (F-CARS) 和双光子激发诱导荧光寿命图像 (FLIM)。我们在不显着损失光、视场的情况下，在非扫描几何结构中使用单根多模光纤实现了这一点，最重要的是，TCSPC 定时保真度。除了展示使用时间门控从 FLIM 图像中分离 CARS 图像的能力外，我们还展示了在活细胞中使用纤维收集 F-CARS 成像的组合多模态上采集 FLIM 成像。
