Hyperspectral imaging with stimulated Raman scattering by chirped femtosecond lasers.
Hyperspectral imaging with stimulated Raman scattering by chirped femtosecond lasers.
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DOI：
10.1021/jp308938t
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发表时间：
2013-04-25
影响因子：
3.3
通讯作者：
Xie, Xiaoliang Sunney
中科院分区：
文献类型：
--
作者：
Fu, Dan;Holtom, Gary;Freudiger, Christian;Zhang, Xu;Xie, Xiaoliang Sunney
拉曼显微术是一种用于研究非均匀体系的定量、无标记和非侵入性光学成像技术。然而，拉曼散射的微弱性显著地限制了拉曼显微术对低时间分辨率和主要静态样品的使用。窄带受激拉曼散射（SRS）显微术的最新发展以减少光谱信息为代价，将基于拉曼的无标记化学成像的灵敏度显著提高了几个数量级。基于光谱聚焦方法，我们提出了一种快速SRS超光谱成像系统，该系统利用啁啾飞秒激光实现快速拉曼光谱采集，同时保持窄带SRS成像的全速和图像质量。我们证明，混合化学物种的定量浓度测定可以实现灵敏度下降到4毫米。成像的目的，整个拉曼光谱数据立方体可以在一分钟内获得。我们发现，哺乳动物细胞SRS高光谱成像揭示了饱和脂质，不饱和脂质，胆固醇和蛋白质的空间不均匀分布。快速光谱和无标记化学成像的结合将使研究生物系统和材料系统的新应用成为可能。
Raman microscopy is a quantitative, label-free and noninvasive optical imaging technique for studying inhomogeneous systems. However, the feebleness of Raman scattering significantly limits the use of Raman microscopy to low time resolutions and primarily static samples. Recent developments in narrowband stimulated Raman Scattering (SRS) microscopy have significantly increased the sensitivity of Raman based label-free chemical imaging by a few orders of magnitude, at the expense of reduced spectroscopic information. Based on a spectral focusing approach, we present a fast SRS hyperspectral imaging system using chirped femtosecond lasers to achieve rapid Raman spectra acquisition while retaining the full speed and image quality of narrowband SRS imaging. We demonstrate that quantitative concentration determination of mixed chemical species can be achieved with sensitivity down to 4 mM. For imaging purposes, an entire Raman spectral data cube can be obtained within a minute. We show that mammalian cells SRS hyperspectral imaging reveals the spatially inhomogeneous distribution of saturated lipids, unsaturated lipids, cholesterol and protein. The combination of fast spectroscopy and label-free chemical imaging will enable new applications in studying biological systems and material systems.
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