Development of a high-speed confocal line scanning FLIM microscope for live cell imaging (Conference Presentation)
Development of a high-speed confocal line scanning FLIM microscope for live cell imaging (Conference Presentation)
复制标题
开发用于活细胞成像的高速共焦线扫描 FLIM 显微镜（会议演示）
DOI：
10.1117/12.2650206
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Poland S
中科院分区：
文献类型：
--
作者：
Poland S
我们报告了一种新型的共焦线扫描显微镜的发展，能够获得视频帧速率TCSPC为基础的FLIM。该系统由一维激光束组成，它与基于1024×16单光子雪崩二极管（SPAD）的线成像CMOS（1）光学共轭，像素间距为23.78 μm，填充因子为49.31%。在线传感器上集成片上直方图有助于采集高达16.5千兆光子计数/秒的数据，使操作速度比我们之前报道的定制高速FLIM平台快66倍。我们将展示其在活细胞成像中的应用，研究PAK蛋白在调节细胞骨架动力学中的作用。
We report the development of a novel confocal line-scanning microscope capable of acquiring video-frame rate TCSPC-based FLIM. The system consists of a one-dimensional laser beam, which is optically conjugated to a 1024×16 single photon avalanche diode(SPAD) based line-imaging CMOS(1), with 23.78 μm pixel pitch at 49.31% fill factor. Incorporation of on-chip histogramming on the line-sensor facilitates the acquisition of up to 16.5 Giga-photon counts/s, enabling operation 66 times faster than our previously reported bespoke high speed FLIM platforms. We will demonstrate its use in live-cell imaging investigating the roles that PAK proteins play in regulation of cytoskeletal dynamics.