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Versatile microporous polymer-based supports for serial macromolecular crystallography
Serial data collection has emerged as a major tool for data collection at state-of-the-art light sources, such as microfocus beamlines at synchrotrons and X-ray free-electron lasers. Challenging targets, characterized by small crystal sizes, weak diffraction and stringent dose limits, benefit most f...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8411977/ https://www.ncbi.nlm.nih.gov/pubmed/34473086 http://dx.doi.org/10.1107/S2059798321007324 |
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author | Martiel, Isabelle Beale, John H. Karpik, Agnieszka Huang, Chia-Ying Vera, Laura Olieric, Natacha Wranik, Maximilian Tsai, Ching-Ju Mühle, Jonas Aurelius, Oskar John, Juliane Högbom, Martin Wang, Meitian Marsh, May Padeste, Celestino |
author_facet | Martiel, Isabelle Beale, John H. Karpik, Agnieszka Huang, Chia-Ying Vera, Laura Olieric, Natacha Wranik, Maximilian Tsai, Ching-Ju Mühle, Jonas Aurelius, Oskar John, Juliane Högbom, Martin Wang, Meitian Marsh, May Padeste, Celestino |
author_sort | Martiel, Isabelle |
collection | PubMed |
description | Serial data collection has emerged as a major tool for data collection at state-of-the-art light sources, such as microfocus beamlines at synchrotrons and X-ray free-electron lasers. Challenging targets, characterized by small crystal sizes, weak diffraction and stringent dose limits, benefit most from these methods. Here, the use of a thin support made of a polymer-based membrane for performing serial data collection or screening experiments is demonstrated. It is shown that these supports are suitable for a wide range of protein crystals suspended in liquids. The supports have also proved to be applicable to challenging cases such as membrane proteins growing in the sponge phase. The sample-deposition method is simple and robust, as well as flexible and adaptable to a variety of cases. It results in an optimally thin specimen providing low background while maintaining minute amounts of mother liquor around the crystals. The 2 × 2 mm area enables the deposition of up to several microlitres of liquid. Imaging and visualization of the crystals are straightforward on the highly transparent membrane. Thanks to their affordable fabrication, these supports have the potential to become an attractive option for serial experiments at synchrotrons and free-electron lasers. |
format | Online Article Text |
id | pubmed-8411977 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-84119772021-09-13 Versatile microporous polymer-based supports for serial macromolecular crystallography Martiel, Isabelle Beale, John H. Karpik, Agnieszka Huang, Chia-Ying Vera, Laura Olieric, Natacha Wranik, Maximilian Tsai, Ching-Ju Mühle, Jonas Aurelius, Oskar John, Juliane Högbom, Martin Wang, Meitian Marsh, May Padeste, Celestino Acta Crystallogr D Struct Biol Research Papers Serial data collection has emerged as a major tool for data collection at state-of-the-art light sources, such as microfocus beamlines at synchrotrons and X-ray free-electron lasers. Challenging targets, characterized by small crystal sizes, weak diffraction and stringent dose limits, benefit most from these methods. Here, the use of a thin support made of a polymer-based membrane for performing serial data collection or screening experiments is demonstrated. It is shown that these supports are suitable for a wide range of protein crystals suspended in liquids. The supports have also proved to be applicable to challenging cases such as membrane proteins growing in the sponge phase. The sample-deposition method is simple and robust, as well as flexible and adaptable to a variety of cases. It results in an optimally thin specimen providing low background while maintaining minute amounts of mother liquor around the crystals. The 2 × 2 mm area enables the deposition of up to several microlitres of liquid. Imaging and visualization of the crystals are straightforward on the highly transparent membrane. Thanks to their affordable fabrication, these supports have the potential to become an attractive option for serial experiments at synchrotrons and free-electron lasers. International Union of Crystallography 2021-08-23 /pmc/articles/PMC8411977/ /pubmed/34473086 http://dx.doi.org/10.1107/S2059798321007324 Text en © Isabelle Martiel et al. 2021 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Martiel, Isabelle Beale, John H. Karpik, Agnieszka Huang, Chia-Ying Vera, Laura Olieric, Natacha Wranik, Maximilian Tsai, Ching-Ju Mühle, Jonas Aurelius, Oskar John, Juliane Högbom, Martin Wang, Meitian Marsh, May Padeste, Celestino Versatile microporous polymer-based supports for serial macromolecular crystallography |
title | Versatile microporous polymer-based supports for serial macromolecular crystallography |
title_full | Versatile microporous polymer-based supports for serial macromolecular crystallography |
title_fullStr | Versatile microporous polymer-based supports for serial macromolecular crystallography |
title_full_unstemmed | Versatile microporous polymer-based supports for serial macromolecular crystallography |
title_short | Versatile microporous polymer-based supports for serial macromolecular crystallography |
title_sort | versatile microporous polymer-based supports for serial macromolecular crystallography |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8411977/ https://www.ncbi.nlm.nih.gov/pubmed/34473086 http://dx.doi.org/10.1107/S2059798321007324 |
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