Abstract

Water-window x-ray microscopy allows two- and three-dimensional (2D and 3D) imaging of intact unstained cells in their cryofixed near-native state with unique contrast and high resolution. Present operational biological water-window microscopes are based at synchrotron facilities, which limits their accessibility and integration with complementary methods. Laboratory-source microscopes have had difficulty addressing relevant biological tasks with proper resolution and contrast due to long exposure times and limited up-time. Here we report on laboratory cryo x-ray microscopy with the exposure time, contrast, and reliability to allow for routine high-spatial resolution 3D imaging of intact cells and cell-cell interactions. Stabilization of the laser-plasma source combined with new optics and sample preparation provide high-resolution cell imaging, both in 2D with ten-second exposures and in 3D with twenty-minute tomography. Examples include monitoring of the distribution of carbon-dense vesicles in starving HEK293T cells and imaging the interaction between natural killer cells and target cells.

Details

Title
Laboratory cryo x-ray microscopy for 3D cell imaging
Author
Fogelqvist, Emelie 1 ; Kördel, Mikael 1 ; Carannante, Valentina 2 ; Önfelt, Björn 3 ; Hertz, Hans M 1 

 Department of Applied Physics, KTH Royal Institute of Technology/Albanova, Stockholm, Sweden 
 Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Solna, Sweden 
 Department of Applied Physics, KTH Royal Institute of Technology/Albanova, Stockholm, Sweden; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Solna, Sweden 
Pages
1-8
Publication year
2017
Publication date
Oct 2017
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2121471269
Copyright
© 2017. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.