光谱时间编码多光子显微术
Spectro-temporal encoded Multiphoton Microscopy
摘要：
双光子显微镜已成为生物和医学研究的宝贵工具，提供高灵敏度，分子特异性，固有的三维亚细胞分辨率和深层组织穿透。然而，就成像速度而言，机械扫描仪仍然将采集速率限制在通常每秒10-100帧。在这里，我们提出了一种高速非线性显微镜实现千赫兹帧速率采用脉冲调制，快速波长扫描激光器和惯性自由光束转向通过角色散。结合高带宽，单光子敏感探测器，我们实现了前所未有的速度每秒8800万像素的荧光寿命记录。我们展示了衍射限制，多模态，双光子荧光和荧光寿命（FLIM），显微镜和成像流式细胞术与数字可重构激光，成像系统和数据采集系统。这些前所未有的速度应该能够实现高速和高通量的图像辅助细胞分选。
英文摘要：
Two-Photon Microscopy has become an invaluable tool for biological and medical research, providing high sensitivity, molecular specificity, inherent three-dimensional sub-cellular resolution and deep tissue penetration. In terms of imaging speeds, however, mechanical scanners still limit the acquisition rates to typically 10-100 frames per second. Here we present a high-speed non-linear microscope achieving kilohertz frame rates by employing pulse-modulated, rapidly wavelength-swept lasers and inertia-free beam steering through angular dispersion. In combination with a high bandwidth, single-photon sensitive detector, we achieve recording of fluorescent lifetimes at unprecedented speeds of 88 million pixels per second. We show diffraction-limited, multi-modal, Two-Photon fluorescence and fluorescence lifetime (FLIM), microscopy and imaging flow cytometry with a digitally reconfigurable laser, imaging system and data acquisition system. These unprecedented speeds should enable high-speed and high-throughput image-assisted cell sorting.
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