Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning
Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning
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DOI：
10.1364/oe.20.008939
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发表时间：
2012-04-09
期刊：
影响因子：
3.8
通讯作者：
Chen, Shean-Jen
中科院分区：
文献类型：
--
作者：
Cheng, Li-Chung;Chang, Chia-Yuan;Chen, Shean-Jen
在本研究中，开发了一种基于时空聚焦的显微镜，提供宽视场多光子激发，以提供快速光学切片图像。该显微镜的主要特点是集成了一个10 kHz重复频率的超快放大器，具有高瞬时峰值功率（在90 fs脉宽下最大400 μ J/脉冲）和一个te冷却，超灵敏的光子探测，电子倍增电荷耦合相机到一个时空聚焦显微镜。这种配置可以产生激发面积大于200 × 100 μ m(2)的多光子图像，帧率大于100 Hz（目前最大200hz）。实时观察到小至0.5 μ m的荧光微珠的布朗运动，横向分辨率小于0.5 μ m，轴向分辨率约为3.5 μ m。此外，鸡肌腱的二次谐波图像表明，所开发的宽视场多光子显微镜可以为生物成像提供高分辨率的z-切面。(C) 2012美国光学学会
In this study, a microscope based on spatiotemporal focusing offering widefield multiphoton excitation has been developed to provide fast optical sectioning images. Key features of this microscope are the integrations of a 10 kHz repetition rate ultrafast amplifier featuring high instantaneous peak power (maximum 400 mu J/pulse at a 90 fs pulse width) and a TE-cooled, ultra-sensitive photon detecting, electron multiplying charge-coupled camera into a spatiotemporal focusing microscope. This configuration can produce multiphoton images with an excitation area larger than 200 x 100 mu m(2) at a frame rate greater than 100 Hz (current maximum of 200 Hz). Brownian motions of fluorescent microbeads as small as 0.5 mu m were observed in real-time with a lateral spatial resolution of less than 0.5 mu m and an axial resolution of approximately 3.5 mu m. Furthermore, second harmonic images of chicken tendons demonstrate that the developed widefield multiphoton microscope can provide high resolution z-sectioning for bioimaging. (C) 2012 Optical Society of America