使用纳秒白光激光源对活细胞进行多模态成像，包括多重相干反斯托克斯拉曼散射（CARS）、三阶和频产生（TSFG）以及双光子激发荧光（TPEF）。

Multimodal imaging of living cells with multiplex coherent anti-stokes raman scattering (CARS), third-order sum frequency generation (TSFG) and two-photon excitation fluorescence (TPEF) using a nanosecond white-light laser source.

作者信息

Segawa Hiroki, Okuno Masanari, Leproux Philippe, Couderc Vincent, Ozawa Takeaki, Kano Hideaki

机构信息

Department of Chemistry, School of Science, The University of Tokyo.

出版信息

Anal Sci. 2015;31(4):299-305. doi: 10.2116/analsci.31.299.

DOI:10.2116/analsci.31.299

PMID:25864673

Abstract

The subnanosecond "white-light laser" source has been applied to multimodal, multiphoton, and multiplex spectroscopic imaging (M(3) spectroscopic imaging) with coherent anti-Stokes Raman scattering (CARS), third-order sum frequency generation (TSFG), and two-photon excitation fluorescence (TPEF). As the proof-of-principle experiment, we performed simultaneous imaging of polystyrene beads with TSFG and TPEF. This technique is then applied to live cell imaging. Mouse L929 fibroblastic cells are clearly visualized by CARS, TSFG, and TPEF processes. M(3) spectroscopic imaging provides various and unique cellular information with different image contrast based on each multiphoton process.

摘要

亚纳秒级“白光激光”源已应用于多模态、多光子和多路复用光谱成像（M(3)光谱成像），包括相干反斯托克斯拉曼散射（CARS）、三阶和频产生（TSFG）以及双光子激发荧光（TPEF）。作为原理验证实验，我们利用TSFG和TPEF对聚苯乙烯微珠进行了同步成像。然后将该技术应用于活细胞成像。通过CARS、TSFG和TPEF过程可以清晰地观察到小鼠L929成纤维细胞。M(3)光谱成像基于每个多光子过程，以不同的图像对比度提供各种独特的细胞信息。