Combination of an inject-and-transfer system for serial femtosecond crystallography
Combination of an inject-and-transfer system for serial femtosecond crystallography
Abstract Serial femtosecond crystallography (SFX) enables the determination of the room-temperature crystal structure of macromolecules without causing radiation damage and provides time-resolved molecular dynamics by pump-probe experiments. In the SFX experiment, the injector-based sample delivery method continuously provides fresh crystals to X-rays, and the fixed-target scanning method can be programmed to move the crystals to the desired location. This study introduces a combination of the inject-and-transfer system (BITS) method for sample delivery for SFX experiments, a hybrid injection, and a fixed-target scanning method. BITS allows solution samples to be reliably deposited on an ultraviolet ozone (UVO)-treaed polyimide films at flow rates as low as 1 nl/min. In application of BITS in SFX experiment, the lysozyme crystal samples were embedded in a viscous lard medium and injected at a 50–100 nl/min flow rate through a syringe needle onto an UVO-treated polyimide film mounted on a fixed-target scan stage. The deposited crystal sample on film were raster scanned to XFEL by motion stage in the horizontal and vertical directions. Using this method, we successfully determined the room-temperature structure of lysozyme at 2.1 ? resolution. This method can be applied to the SFX experiments.
Lee Keondo、Baek Sangwon、Park Jaehyun、Park Sehan、Chung Wan Kyun、Nam Ki Hyun、Lee Jong-Lam、Cho Yunje、Kim Jihan
Department of Mechanical Engineering, Pohang University of Science and TechnologyDepartment of Materials Science and Engineering, Pohang University of Science and TechnologyPohang Accelerator Laboratory, Pohang University of Science and Technology||Department of Chemical Engineering, Pohang University of Science and TechnologyPohang Accelerator Laboratory, Pohang University of Science and TechnologyDepartment of Mechanical Engineering, Pohang University of Science and TechnologyDepartment of Life Science, Pohang University of Science and Technology||POSTECH Biotech Center, Pohang University of Science and TechnologyDepartment of Materials Science and Engineering, Pohang University of Science and TechnologyDepartment of Life Science, Pohang University of Science and TechnologyDepartment of Life Science, Pohang University of Science and Technology
晶体学生物化学生物物理学
serial crystallographysample deliveryinjectionfixed-target scanningviscous medium
Lee Keondo,Baek Sangwon,Park Jaehyun,Park Sehan,Chung Wan Kyun,Nam Ki Hyun,Lee Jong-Lam,Cho Yunje,Kim Jihan.Combination of an inject-and-transfer system for serial femtosecond crystallography[EB/OL].(2025-03-28)[2025-08-02].https://www.biorxiv.org/content/10.1101/2022.02.08.479470.点此复制
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