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Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography
The serial crystallography (SX) technique enables the determination of the room-temperature structure of a macromolecule while causing minimal radiation damage, as well as the visualization of the molecular dynamics by time-resolved studies. The fixed-target (FT) scanning approach is one method for...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8222285/ https://www.ncbi.nlm.nih.gov/pubmed/34162965 http://dx.doi.org/10.1038/s41598-021-92687-x |
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author | Nam, Ki Hyun Kim, Jihan Cho, Yunje |
author_facet | Nam, Ki Hyun Kim, Jihan Cho, Yunje |
author_sort | Nam, Ki Hyun |
collection | PubMed |
description | The serial crystallography (SX) technique enables the determination of the room-temperature structure of a macromolecule while causing minimal radiation damage, as well as the visualization of the molecular dynamics by time-resolved studies. The fixed-target (FT) scanning approach is one method for SX sample delivery that minimizes sample consumption and minimizes physical damage to crystals during data collection. Settling of the crystals on the sample holder in random orientation is important for complete three dimensional data collection. To increase the random orientation of crystals on the sample holder, we developed a polyimide mesh-based sample holder with irregular crystal mounting holes for FT-SX. The polyimide mesh was fabricated using a picosecond laser. Each hole in the polyimide mesh has irregularly shaped holes because of laser thermal damage, which may cause more crystals to settle at random orientations compared to regular shaped sample holders. A crystal sample was spread onto a polyimide-mesh, and a polyimide film was added to both sides to prevent dehydration. Using this sample holder, FT-SX was performed at synchrotron and determined the room-temperature lysozyme structure at 1.65 Å. The polyimide mesh with irregularly shaped holes will allow for expanded applications in sample delivery for FT-SX experiments. |
format | Online Article Text |
id | pubmed-8222285 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-82222852021-06-24 Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography Nam, Ki Hyun Kim, Jihan Cho, Yunje Sci Rep Article The serial crystallography (SX) technique enables the determination of the room-temperature structure of a macromolecule while causing minimal radiation damage, as well as the visualization of the molecular dynamics by time-resolved studies. The fixed-target (FT) scanning approach is one method for SX sample delivery that minimizes sample consumption and minimizes physical damage to crystals during data collection. Settling of the crystals on the sample holder in random orientation is important for complete three dimensional data collection. To increase the random orientation of crystals on the sample holder, we developed a polyimide mesh-based sample holder with irregular crystal mounting holes for FT-SX. The polyimide mesh was fabricated using a picosecond laser. Each hole in the polyimide mesh has irregularly shaped holes because of laser thermal damage, which may cause more crystals to settle at random orientations compared to regular shaped sample holders. A crystal sample was spread onto a polyimide-mesh, and a polyimide film was added to both sides to prevent dehydration. Using this sample holder, FT-SX was performed at synchrotron and determined the room-temperature lysozyme structure at 1.65 Å. The polyimide mesh with irregularly shaped holes will allow for expanded applications in sample delivery for FT-SX experiments. Nature Publishing Group UK 2021-06-23 /pmc/articles/PMC8222285/ /pubmed/34162965 http://dx.doi.org/10.1038/s41598-021-92687-x Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Nam, Ki Hyun Kim, Jihan Cho, Yunje Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography |
title | Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography |
title_full | Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography |
title_fullStr | Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography |
title_full_unstemmed | Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography |
title_short | Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography |
title_sort | polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8222285/ https://www.ncbi.nlm.nih.gov/pubmed/34162965 http://dx.doi.org/10.1038/s41598-021-92687-x |
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