Cargando…
X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning
X-ray microscopy can be used to image whole, unsectioned cells in their native hydrated state. It complements the higher resolution of electron microscopy for submicrometer thick specimens, and the molecule-specific imaging capabilites of fluorescence light microscopy. We describe here the first use...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5428657/ https://www.ncbi.nlm.nih.gov/pubmed/28348401 http://dx.doi.org/10.1038/s41598-017-00569-y |
_version_ | 1783235870623531008 |
---|---|
author | Deng, Junjing Vine, David J. Chen, Si Jin, Qiaoling Nashed, Youssef S. G. Peterka, Tom Vogt, Stefan Jacobsen, Chris |
author_facet | Deng, Junjing Vine, David J. Chen, Si Jin, Qiaoling Nashed, Youssef S. G. Peterka, Tom Vogt, Stefan Jacobsen, Chris |
author_sort | Deng, Junjing |
collection | PubMed |
description | X-ray microscopy can be used to image whole, unsectioned cells in their native hydrated state. It complements the higher resolution of electron microscopy for submicrometer thick specimens, and the molecule-specific imaging capabilites of fluorescence light microscopy. We describe here the first use of fast, continuous x-ray scanning of frozen hydrated cells for simultaneous sub-20 nm resolution ptychographic transmission imaging with high contrast, and sub-100 nm resolution deconvolved x-ray fluorescence imaging of diffusible and bound ions at native concentrations, without the need to add specific labels. By working with cells that have been rapidly frozen without the use of chemical fixatives, and imaging them under cryogenic conditions, we are able to obtain images with well preserved structural and chemical composition, and sufficient stability against radiation damage to allow for multiple images to be obtained with no observable change. |
format | Online Article Text |
id | pubmed-5428657 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-54286572017-05-15 X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning Deng, Junjing Vine, David J. Chen, Si Jin, Qiaoling Nashed, Youssef S. G. Peterka, Tom Vogt, Stefan Jacobsen, Chris Sci Rep Article X-ray microscopy can be used to image whole, unsectioned cells in their native hydrated state. It complements the higher resolution of electron microscopy for submicrometer thick specimens, and the molecule-specific imaging capabilites of fluorescence light microscopy. We describe here the first use of fast, continuous x-ray scanning of frozen hydrated cells for simultaneous sub-20 nm resolution ptychographic transmission imaging with high contrast, and sub-100 nm resolution deconvolved x-ray fluorescence imaging of diffusible and bound ions at native concentrations, without the need to add specific labels. By working with cells that have been rapidly frozen without the use of chemical fixatives, and imaging them under cryogenic conditions, we are able to obtain images with well preserved structural and chemical composition, and sufficient stability against radiation damage to allow for multiple images to be obtained with no observable change. Nature Publishing Group UK 2017-03-27 /pmc/articles/PMC5428657/ /pubmed/28348401 http://dx.doi.org/10.1038/s41598-017-00569-y Text en © The Author(s) 2017 This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Deng, Junjing Vine, David J. Chen, Si Jin, Qiaoling Nashed, Youssef S. G. Peterka, Tom Vogt, Stefan Jacobsen, Chris X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning |
title | X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning |
title_full | X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning |
title_fullStr | X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning |
title_full_unstemmed | X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning |
title_short | X-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning |
title_sort | x-ray ptychographic and fluorescence microscopy of frozen-hydrated cells using continuous scanning |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5428657/ https://www.ncbi.nlm.nih.gov/pubmed/28348401 http://dx.doi.org/10.1038/s41598-017-00569-y |
work_keys_str_mv | AT dengjunjing xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning AT vinedavidj xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning AT chensi xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning AT jinqiaoling xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning AT nashedyoussefsg xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning AT peterkatom xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning AT vogtstefan xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning AT jacobsenchris xrayptychographicandfluorescencemicroscopyoffrozenhydratedcellsusingcontinuousscanning |