Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, 226-8503, Japan.
Curr Pharm Biotechnol. 2013;14(2):159-66.
For many years, spatial resolution is the most critical problem in IR microspectroscopy. This is because the spatial resolution of a conventional infrared microscope is restricted by the diffraction limit, which is almost the same as the wavelength of IR light, ranging from 2.5 to 25 μm. In the recent years, we have developed two novel types of far-field IR super-resolution microscopes using 2-color laser spectroscopies, those are transient fluorescence detected IR (TFD-IR) spectroscopy and vibrational sum-frequency generation (VSFG) spectroscopy. In these ways, because both transient fluorescence and VSFG signal have a wavelength in the visible region, the image is observed at the resolution of visible light, which is about 10 times smaller than that of IR light (that is, IR super-resolution). By using these techniques, we can map the specific IR absorption band with sub-micrometer spatial resolution, visualization of the molecular structure and reaction dynamics in a non-uniform environment such as a cell becomes a possibility. In the present reviews, we introduce our novel IR super-resolution microspectroscopy and its application to single cells in detail.
多年来，空间分辨率一直是红外光谱学中最关键的问题。这是因为传统红外显微镜的空间分辨率受到衍射极限的限制，其大小几乎与红外光的波长相同，范围从 2.5 到 25 μm。近年来，我们使用双色激光光谱学开发了两种新型远场红外超分辨率显微镜，即瞬态荧光检测红外（TFD-IR）光谱和振动和频产生（VSFG）光谱。在这些方法中，由于瞬态荧光和 VSFG 信号都具有可见光区域的波长，因此可以在可见光的分辨率下观察图像，这比红外光的分辨率小大约 10 倍（即红外超分辨率）。通过使用这些技术，我们可以以亚微米空间分辨率绘制特定的红外吸收带，使在非均匀环境（如细胞）中可视化分子结构和反应动力学成为可能。在本综述中，我们详细介绍了我们的新型红外超分辨率光谱学及其在单细胞中的应用。
