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Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals
A fast continuous grid scan protocol has been incorporated into the Swiss Light Source (SLS) data acquisition and analysis software suite on the macromolecular crystallography (MX) beamlines. Its combination with fast readout single-photon counting hybrid pixel array detectors (PILATUS and EIGER) al...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4886984/ https://www.ncbi.nlm.nih.gov/pubmed/27275141 http://dx.doi.org/10.1107/S1600576716006233 |
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author | Wojdyla, Justyna Aleksandra Panepucci, Ezequiel Martiel, Isabelle Ebner, Simon Huang, Chia-Ying Caffrey, Martin Bunk, Oliver Wang, Meitian |
author_facet | Wojdyla, Justyna Aleksandra Panepucci, Ezequiel Martiel, Isabelle Ebner, Simon Huang, Chia-Ying Caffrey, Martin Bunk, Oliver Wang, Meitian |
author_sort | Wojdyla, Justyna Aleksandra |
collection | PubMed |
description | A fast continuous grid scan protocol has been incorporated into the Swiss Light Source (SLS) data acquisition and analysis software suite on the macromolecular crystallography (MX) beamlines. Its combination with fast readout single-photon counting hybrid pixel array detectors (PILATUS and EIGER) allows for diffraction-based identification of crystal diffraction hotspots and the location and centering of membrane protein microcrystals in the lipid cubic phase (LCP) in in meso in situ serial crystallography plates and silicon nitride supports. Diffraction-based continuous grid scans with both still and oscillation images are supported. Examples that include a grid scan of a large (50 nl) LCP bolus and analysis of the resulting diffraction images are presented. Scanning transmission X-ray microscopy (STXM) complements and benefits from fast grid scanning. STXM has been demonstrated at the SLS beamline X06SA for near-zero-dose detection of protein crystals mounted on different types of sample supports at room and cryogenic temperatures. Flash-cooled crystals in nylon loops were successfully identified in differential and integrated phase images. Crystals of just 10 µm thickness were visible in integrated phase images using data collected with the EIGER detector. STXM offers a truly low-dose method for locating crystals on solid supports prior to diffraction data collection at both synchrotron microfocusing and free-electron laser X-ray facilities. |
format | Online Article Text |
id | pubmed-4886984 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-48869842016-06-06 Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals Wojdyla, Justyna Aleksandra Panepucci, Ezequiel Martiel, Isabelle Ebner, Simon Huang, Chia-Ying Caffrey, Martin Bunk, Oliver Wang, Meitian J Appl Crystallogr Research Papers A fast continuous grid scan protocol has been incorporated into the Swiss Light Source (SLS) data acquisition and analysis software suite on the macromolecular crystallography (MX) beamlines. Its combination with fast readout single-photon counting hybrid pixel array detectors (PILATUS and EIGER) allows for diffraction-based identification of crystal diffraction hotspots and the location and centering of membrane protein microcrystals in the lipid cubic phase (LCP) in in meso in situ serial crystallography plates and silicon nitride supports. Diffraction-based continuous grid scans with both still and oscillation images are supported. Examples that include a grid scan of a large (50 nl) LCP bolus and analysis of the resulting diffraction images are presented. Scanning transmission X-ray microscopy (STXM) complements and benefits from fast grid scanning. STXM has been demonstrated at the SLS beamline X06SA for near-zero-dose detection of protein crystals mounted on different types of sample supports at room and cryogenic temperatures. Flash-cooled crystals in nylon loops were successfully identified in differential and integrated phase images. Crystals of just 10 µm thickness were visible in integrated phase images using data collected with the EIGER detector. STXM offers a truly low-dose method for locating crystals on solid supports prior to diffraction data collection at both synchrotron microfocusing and free-electron laser X-ray facilities. International Union of Crystallography 2016-05-16 /pmc/articles/PMC4886984/ /pubmed/27275141 http://dx.doi.org/10.1107/S1600576716006233 Text en © Justyna Aleksandra Wojdyla et al. 2016 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Wojdyla, Justyna Aleksandra Panepucci, Ezequiel Martiel, Isabelle Ebner, Simon Huang, Chia-Ying Caffrey, Martin Bunk, Oliver Wang, Meitian Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals |
title | Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals |
title_full | Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals |
title_fullStr | Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals |
title_full_unstemmed | Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals |
title_short | Fast two-dimensional grid and transmission X-ray microscopy scanning methods for visualizing and characterizing protein crystals |
title_sort | fast two-dimensional grid and transmission x-ray microscopy scanning methods for visualizing and characterizing protein crystals |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4886984/ https://www.ncbi.nlm.nih.gov/pubmed/27275141 http://dx.doi.org/10.1107/S1600576716006233 |
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