Analytic 3D imaging of mammalian nucleus at nanoscale using coherent x-rays and optical fluorescence microscopy

Changyong Song; Masatoshi Takagi; Jaehyun Park; Rui Xu; Marcus Gallagher-Jones; Naoko Imamoto; Tetsuya Ishikawa

doi:10.1016/j.bpj.2014.07.028

Analytic 3D imaging of mammalian nucleus at nanoscale using coherent x-rays and optical fluorescence microscopy

Biophys J. 2014 Sep 2;107(5):1074-1081. doi: 10.1016/j.bpj.2014.07.028.

Authors

Changyong Song¹, Masatoshi Takagi², Jaehyun Park³, Rui Xu⁴, Marcus Gallagher-Jones⁵, Naoko Imamoto², Tetsuya Ishikawa³

Affiliations

¹ RIKEN SPring-8 Center, Sayo, Hyogo, Japan. Electronic address: [email protected].
² RIKEN, 2-1 Hirosawa, Wako-shi, Saitama, Japan.
³ RIKEN SPring-8 Center, Sayo, Hyogo, Japan.
⁴ RIKEN SPring-8 Center, Sayo, Hyogo, Japan; Department of Physics and Astronomy, University of California, Los Angeles, California.
⁵ RIKEN SPring-8 Center, Sayo, Hyogo, Japan; Institute of Integrative Biology, University of Liverpool, Liverpool, UK.

Abstract

Despite the notable progress that has been made with nano-bio imaging probes, quantitative nanoscale imaging of multistructured specimens such as mammalian cells remains challenging due to their inherent structural complexity. Here, we successfully performed three-dimensional (3D) imaging of mammalian nuclei by combining coherent x-ray diffraction microscopy, explicitly visualizing nuclear substructures at several tens of nanometer resolution, and optical fluorescence microscopy, cross confirming the substructures with immunostaining. This demonstrates the successful application of coherent x-rays to obtain the 3D ultrastructure of mammalian nuclei and establishes a solid route to nanoscale imaging of complex specimens.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Cell Nucleus / ultrastructure*
Fluorescent Antibody Technique
Imaging, Three-Dimensional / methods*
Mice
Microscopy, Fluorescence / methods*
Optical Imaging / methods*
X-Ray Diffraction / methods*