Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Nam, Ki Hyun, Kim, Jihan, Cho, Yunje
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
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
_version_ 1783711462220365824
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
work_keys_str_mv AT namkihyun polyimidemeshbasedsampleholderwithirregularcrystalmountingholesforfixedtargetserialcrystallography
AT kimjihan polyimidemeshbasedsampleholderwithirregularcrystalmountingholesforfixedtargetserialcrystallography
AT choyunje polyimidemeshbasedsampleholderwithirregularcrystalmountingholesforfixedtargetserialcrystallography