ja7b03899_si_001.pdf (10.13 MB)
Three-Dimensional Localization of an Individual Fluorescent Molecule with Angstrom Precision
journal contribution
posted on 2017-06-22, 21:04 authored by Taku Furubayashi, Kazuya Motohashi, Keisuke Wakao, Tsuyoshi Matsuda, Isao Kii, Takamitsu Hosoya, Nobuhiro Hayashi, Mahito Sadaie, Fuyuki Ishikawa, Michio Matsushita, Satoru FujiyoshiAmong
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 106 fluorescence
photons from the molecule were accumulated in 5 min.