Ultrafast confocal fluorescence microscopy beyond the fluorescence lifetime limit
Ultrafast confocal fluorescence microscopy beyond the fluorescence lifetime limit
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DOI：
10.1364/optica.5.000117
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发表时间：
2018-02-20
期刊：
影响因子：
10.4
通讯作者：
Goda, Keisuke
中科院分区：
文献类型：
--
作者：
Mikami, Hideharu;Harmon, Jeffrey;Goda, Keisuke
激光扫描共聚焦荧光显微镜因其高空间分辨率而成为生物医学研究中不可缺少的工具。它的时间分辨率同样重要，但对于许多应用来说仍然不够。在这里，我们提出了一个共聚焦荧光显微镜，第一次，我们的知识，超过了最高可能的帧速率仅受荧光团的荧光寿命（通常是几到几纳秒）。这种显微镜是通过集成宽带，空间分布，双频梳或空间双梳和正交幅度调制，以优化频谱效率到频分复用与单像素光电检测信号集成。具体来说，我们展示了共焦荧光显微镜在创纪录的高帧速率为16,000帧/秒。为了展示其广泛的生物医学用途，我们使用显微镜以104体积/s的速度展示细胞动力学的3D体积共聚焦荧光显微镜，并以高达2 m/s的速度展示血液和微藻细胞的共聚焦荧光成像流式细胞术。(c)根据OSA开放获取出版协议的条款，2018年美国光学学会
Laser-scanning confocal fluorescence microscopy is an indispensable tool for biomedical research by virtue of its high spatial resolution. Its temporal resolution is equally important, but is still inadequate for many applications. Here we present a confocal fluorescence microscope that, for the first time to our knowledge, surpasses the highest possible frame rate constrained only by the fluorescence lifetime of fluorophores (typically a few to several nanoseconds). This microscope is enabled by integrating a broadband, spatially distributed, dual-frequency comb or spatial dual-comb and quadrature amplitude modulation for optimizing spectral efficiency into frequency-division multiplexing with single-pixel photodetection for signal integration. Specifically, we demonstrate confocal fluorescence microscopy at a record high frame rate of 16,000 frames/s. To show its broad biomedical utility, we use the microscope to demonstrate 3D volumetric confocal fluorescence microscopy of cellular dynamics at 104 volumes/s and confocal fluorescence imaging flow cytometry of hematological and microalgal cells at up to 2 m/s. (c) 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement