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.

中文翻译：

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超快速共聚焦荧光显微镜超过了荧光寿命极限

激光扫描共聚焦荧光显微镜凭借其高空间分辨率是生物医学研究必不可少的工具。它的时间分辨率同样重要，但对于许多应用程序仍然不够。在这里，我们介绍了一种共聚焦荧光显微镜，据我们所知，它首次超过了仅受荧光团的荧光寿命（通常为几纳秒）约束的最高帧速率。通过将宽带，空间分布的双频梳或空间双梳和正交幅度调制集成到一起以实现信号效率的单像素光电检测的频分多路复用，以优化光谱效率，从而使该显微镜成为可能。具体来说，我们证明了共聚焦荧光显微镜在创纪录的高帧频为16时，000帧/秒 为了显示其广泛的生物医学实用性，我们使用显微镜来演示3D体积共聚焦荧光显微镜下细胞动力学的104体积/秒和共聚焦荧光成像流式细胞术的血液学和微藻类细胞高达2 m / s。