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A simple goniometer-compatible flow cell for serial synchrotron X-ray crystallography

Swagatha Ghosh, Doris Zoric, Peter Dahl,Monika Bjelcic,Jonatan Johannesson,Emil Sandelin,Per Borjesson, Alexander Bjorling,Analia Banacore,Petra Edlund, Oskar Aurelius,Mirko Milas, Jie Nan, Anastasya Shilova,Ana Gonzalez,Uwe Mueller, Gisela Branden,Richard Neutze

Journal of applied crystallography(2023)

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Abstract
Serial femtosecond crystallography was initially developed for roomtemperature X-ray diffraction studies of macromolecules at X-ray free electron lasers. When combined with tools that initiate biological reactions within microcrystals, time-resolved serial crystallography allows the study of structural changes that occur during an enzyme catalytic reaction. Serial synchrotron X-ray crystallography (SSX), which extends serial crystallography methods to synchrotron radiation sources, is expanding the scientific community using serial diffraction methods. This report presents a simple flow cell that can be used to deliver microcrystals across an X-ray beam during SSX studies. This device consists of an X-ray transparent glass capillary mounted on a goniometercompatible 3D-printed support and is connected to a syringe pump via lightweight tubing. This flow cell is easily mounted and aligned, and it is disposable so can be rapidly replaced when blocked. This system was demonstrated by collecting SSX data at MAX IV Laboratory from microcrystals of the integral membrane protein cytochrome c oxidase from Thermus thermophilus, from which an X-ray structure was determined to 2.12 angstrom resolution. This simple SSX platform may help to lower entry barriers for non-expert users of SSX.
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Key words
serial synchrotron X-ray crystallography,macromolecular crystallography,cytochrome c oxidase,goniometer-compatible flow cells
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