Won Youngjae, Moon Sucbei, Yang Wenzhong, Kim Donguk, Han Won-Taek, Kim Dug Young
Department of Information and Communications, Gwangju Institute of Science and Technology, 1 Oryong-dong, Buk-gu, Gwangju 500-712, South Korea.
Opt Express. 2011 Feb 14;19(4):3396-405. doi: 10.1364/OE.19.003396.
We demonstrate a high-speed confocal fluorescence lifetime imaging microscopy (FLIM) whose accuracy and photon economy are as good as that of a time-correlated single photon counting (TCSPC). It is based on a new lifetime determination scheme, the analog mean delay (AMD) method. Due to the technical advantages of multiple fluorescence photon detection capability, accurate lifetime determination scheme and high photon detection efficiency, the AMD method can be the most effective method for high-speed confocal FLIM. The feasibility of real-time confocal FLIM with the AMD method has been demonstrated by observing the dynamic reaction of calcium channels in a RBL-2H3 cell with respect to 4αPDD stimulus. We have achieved the photon detection rate of 125 times faster than a conventional TCSPC based system in this experiment.
我们展示了一种高速共聚焦荧光寿命成像显微镜（FLIM），其精度和光子经济性与时间相关单光子计数（TCSPC）相当。它基于一种新的寿命测定方案，即模拟平均延迟（AMD）方法。由于具有多荧光光子检测能力、精确的寿命测定方案和高光子检测效率等技术优势，AMD方法可能是用于高速共聚焦FLIM的最有效方法。通过观察RBL - 2H3细胞中钙通道对4αPDD刺激的动态反应，证明了采用AMD方法进行实时共聚焦FLIM的可行性。在本实验中，我们实现了比传统基于TCSPC的系统快125倍的光子检测速率。
