Department of Chemistry, School of Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan, 113-0033.
Opt Lett. 2010 Dec 15;35(24):4096-8. doi: 10.1364/OL.35.004096.
We have developed a multifocus confocal Raman microspectroscopic system for the fast multimode vibrational imaging of living cells. It consists of an inverted microscope equipped with a microlens array, a pinhole array, a fiber bundle, and a multichannel Raman spectrometer. Forty-eight Raman spectra from 48 foci under the microscope are simultaneously obtained by using multifocus excitation and image-compression techniques. The multifocus confocal configuration suppresses the background generated from the cover glass and the cell culturing medium so that high-contrast images are obtainable with a short accumulation time. The system enables us to obtain multimode (10 different vibrational modes) vibrational images of living cells in tens of seconds with only 1 mW laser power at one focal point. This image acquisition time is more than 10 times faster than that in conventional single-focus Raman microspectroscopy.
我们开发了一种用于快速多模式活细胞振动成像的多焦点共焦 Raman 微光谱系统。它由一个倒置显微镜组成，该显微镜配备了微透镜阵列、针孔阵列、光纤束和多通道 Raman 光谱仪。通过使用多焦点激发和图像压缩技术，从显微镜下的 48 个焦点同时获得 48 个 Raman 光谱。多焦点共焦配置抑制了来自盖玻片和细胞培养液的背景，从而可以在短的积累时间内获得高对比度的图像。该系统使我们能够在一个焦点处仅用 1 mW 激光功率在几十秒内获得活细胞的多模（10 种不同的振动模式）振动图像。与传统的单焦点 Raman 微光谱相比，这种图像采集时间快了 10 多倍。
