Pathophysiological and Health Science Team, Imaging Platform and Innovation Group, Division of Bio-Function Dynamics Imaging, RIKEN Center for Life Science Technologies , Chuo, Kobe, 650-0047, Japan.
Laboratory of Chemical Bioscience, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University , Chiyoda, Tokyo, 101-0062, Japan.
J Am Chem Soc. 2017 Jul 5;139(26):8990-8994. doi: 10.1021/jacs.7b03899. Epub 2017 Jun 23.
Among imaging techniques, fluorescence microscopy is a unique method to noninvasively image individual molecules in whole cells. If the three-dimensional spatial precision is improved to the angstrom level, various molecular arrangements in the cell can be visualized on an individual basis. We have developed a cryogenic reflecting microscope with a numerical aperture of 0.99 and an imaging stability of 0.05 nm in standard deviation at a temperature of 1.8 K. The key optics to realize the cryogenic performances is the reflecting objective developed by our laboratory. With this cryogenic microscope, an individual fluorescent molecule (ATTO647N) at 1.8 K was localized with standard errors of 0.53 nm (x), 0.31 nm (y), and 0.90 nm (z) when 10 fluorescence photons from the molecule were accumulated in 5 min.
在成像技术中，荧光显微镜是一种独特的方法，可以非侵入性地对整个细胞中的单个分子进行成像。如果将三维空间精度提高到埃级，就可以在单个基础上可视化细胞中的各种分子排列。我们开发了一种具有 0.99 的数值孔径和在 1.8 K 温度下标准差为 0.05nm 的成像稳定性的低温反射显微镜。实现低温性能的关键光学元件是我们实验室开发的反射物镜。使用这种低温显微镜，当在 5 分钟内累积来自分子的 10 个荧光光子时，1.8K 下的单个荧光分子（ATTO647N）的定位标准误差为 0.53nm（x）、0.31nm（y）和 0.90nm（z）。
