Murakami Yusuke, Ando Masahiro, Imamura Ayako, Oketani Ryosuke, Leproux Philippe, Honjoh Sakiko, Kano Hideaki
Ph.D. Program in Humanics, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan.
International Institute for Integrative Sleep Medicine (WPI-IIIS), 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
Chem Biomed Imaging. 2024 Jul 25;2(10):689-697. doi: 10.1021/cbmi.4c00034. eCollection 2024 Oct 28.
The Raman fingerprint spectral region provides abundant structural information on molecules. However, analyzing vibrational images within this region using coherent Raman imaging remains challenging due to the small Raman cross section and congested spectral features. In this study, we combined ultrabroadband coherent anti-Stokes Raman scattering (CARS) microspectroscopy across the spectral range of 500-4000 cm with multivariate curve resolution-alternating least-squares (MCR-ALS) to reveal hidden Raman bands in the fingerprint region. Applying this method to mouse brain tissue, we extracted information on cholesterol and collagen, leveraging their distinctive molecular signatures, as well as on key molecules such as lipids, proteins, water, and nucleic acids. Moreover, the simultaneous detection of second harmonic generation facilitated label-free visualization of organelles, including arachnoid membrane and Rootletin filaments.
拉曼指纹光谱区域提供了关于分子的丰富结构信息。然而，由于拉曼散射截面小且光谱特征拥挤，使用相干拉曼成像分析该区域内的振动图像仍然具有挑战性。在本研究中，我们将500 - 4000 cm光谱范围内的超宽带相干反斯托克斯拉曼散射（CARS）显微光谱与多元曲线分辨交替最小二乘法（MCR - ALS）相结合，以揭示指纹区域中隐藏的拉曼谱带。将该方法应用于小鼠脑组织，我们利用胆固醇、胶原蛋白以及脂质、蛋白质、水和核酸等关键分子独特的分子特征提取了相关信息。此外，二次谐波产生的同时检测有助于对包括蛛网膜和根蛋白细丝在内的细胞器进行无标记可视化。
