Department of Electrical Engineering and Information Systems, The University of Tokyo, Tokyo 113-8656, Japan.
Department of Applied Biological Science, Tokyo University of Science, Noda 278-8510, Japan.
Analyst. 2021 Feb 22;146(4):1234-1238. doi: 10.1039/d0an02181d.
Recent development of stimulated Raman scattering (SRS) microscopy allows for label-free biological imaging with chemical specificity based on molecular-vibrational signatures. In particular, hyperspectral SRS imaging can acquire a molecular-vibrational spectrum at each pixel, allowing us not only to investigate the spectral difference of various biological molecules but also to discriminate different constituents based on their spectral difference. However, the number of constituents discriminated in previous label-free SRS imaging was limited to four because of the subtleness of spectral difference. Here, we report hyperspectral SRS imaging of plant tissues including leaves of Camellia japonica, roots of Arabidopsis thaliana, and thalli of a liverwort Marchantia polymorpha L. We show that SRS can discriminate as many as six components in Marchantia polymorpha L. without labeling. Our results demonstrate the effectiveness of hyperspectral SRS imaging as a tool for label-free multicolour imaging analysis of various biomolecules in plant tissues.
最近，受激拉曼散射（SRS）显微镜的发展允许基于分子振动特征进行无标记的具有化学特异性的生物成像。特别是，高光谱 SRS 成像可以在每个像素处获取分子振动光谱，这不仅允许我们研究各种生物分子的光谱差异，还允许我们基于它们的光谱差异来区分不同的成分。然而，由于光谱差异的细微性，以前的无标记 SRS 成像中能够区分的成分数量被限制为四种。在这里，我们报告了包括茶树叶片、拟南芥根和地钱叶状体在内的植物组织的高光谱 SRS 成像。我们表明，SRS 可以在无需标记的情况下区分多达六种地钱叶状体成分。我们的结果证明了高光谱 SRS 成像作为一种工具的有效性，用于对植物组织中的各种生物分子进行无标记的多色成像分析。
