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Laboratory cryo x-ray microscopy for 3D cell imaging

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...

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Autores principales: Fogelqvist, Emelie, Kördel, Mikael, Carannante, Valentina, Önfelt, Björn, Hertz, Hans M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5647409/
https://www.ncbi.nlm.nih.gov/pubmed/29044158
http://dx.doi.org/10.1038/s41598-017-13538-2
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author Fogelqvist, Emelie
Kördel, Mikael
Carannante, Valentina
Önfelt, Björn
Hertz, Hans M.
author_facet Fogelqvist, Emelie
Kördel, Mikael
Carannante, Valentina
Önfelt, Björn
Hertz, Hans M.
author_sort Fogelqvist, Emelie
collection PubMed
description 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.
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spelling pubmed-56474092017-10-26 Laboratory cryo x-ray microscopy for 3D cell imaging Fogelqvist, Emelie Kördel, Mikael Carannante, Valentina Önfelt, Björn Hertz, Hans M. Sci Rep Article 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. Nature Publishing Group UK 2017-10-18 /pmc/articles/PMC5647409/ /pubmed/29044158 http://dx.doi.org/10.1038/s41598-017-13538-2 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Fogelqvist, Emelie
Kördel, Mikael
Carannante, Valentina
Önfelt, Björn
Hertz, Hans M.
Laboratory cryo x-ray microscopy for 3D cell imaging
title Laboratory cryo x-ray microscopy for 3D cell imaging
title_full Laboratory cryo x-ray microscopy for 3D cell imaging
title_fullStr Laboratory cryo x-ray microscopy for 3D cell imaging
title_full_unstemmed Laboratory cryo x-ray microscopy for 3D cell imaging
title_short Laboratory cryo x-ray microscopy for 3D cell imaging
title_sort laboratory cryo x-ray microscopy for 3d cell imaging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5647409/
https://www.ncbi.nlm.nih.gov/pubmed/29044158
http://dx.doi.org/10.1038/s41598-017-13538-2
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