Fukuta Masahiro, Kanamori Satoshi, Furukawa Taichi, Nawa Yasunori, Inami Wataru, Lin Sheng, Kawata Yoshimasa, Terakawa Susumu
Graduate School of Science and Technology, Shizuoka University, 3-5-1 Johoku, Naka, Hamamatsu 432-8561, Japan.
Institute of NanoScience Design, Osaka University, 1-3, Machikaneyamacho, Toyonaka 560-0043, Japan.
Sci Rep. 2015 Nov 3;5:16068. doi: 10.1038/srep16068.
Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.
光学显微镜是细胞功能分析的有效工具，因为在水或大气条件下，可以在活状态下对生物细胞进行无损、非侵入性观察。诸如相差显微镜等无标记光学成像技术已被用于分析许多细胞功能，它是生物科学领域的一项关键技术。然而，光的衍射极限使得在无标记的活细胞中对纳米结构成像变得困难，例如内质网、高尔基体和蛋白质的定位。在这里，我们展示了使用电子束激发辅助光学（EXA）显微镜对无标记细胞进行高空间分辨率的动态成像。我们以优于100纳米的空间分辨率和大约10的信噪比观察了活细胞中细胞核和纳米级颗粒的动态运动。我们的结果有助于细胞功能分析的发展，并开辟新的生物科学应用。
