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Fixed-Target Serial Synchrotron Crystallography Using Nylon Mesh and Enclosed Film-Based Sample Holder

1
Pohang Accelerator Laboratory, Pohang, Gyeongbuk 37673, Korea
2
Department of Life Science, Pohang University of Science and Technology, Pohang, Gyeongbuk 37673, Korea
*
Author to whom correspondence should be addressed.
Crystals 2020, 10(9), 803; https://doi.org/10.3390/cryst10090803
Received: 28 August 2020 / Revised: 8 September 2020 / Accepted: 9 September 2020 / Published: 10 September 2020
(This article belongs to the Special Issue Approach of Serial Crystallography)
Serial crystallography (SX) technique using synchrotron X-ray allows the visualization of room-temperature crystal structures with low-dose data collection as well as time-resolved molecular dynamics. In an SX experiment, delivery of numerous crystals for X-ray interaction, in a serial manner, is very important. Fixed-target scanning approach has the advantage of dramatically minimizing sample consumption as well as any physical damage to crystal sample, compared to other sample delivery methods. Here, we introduce the simple approach of fixed-target serial synchrotron crystallography (FT-SSX) using nylon mesh and enclosed film (NAM)-based sample holder. The NAM-based sample holder consisted of X-ray-transparent nylon-mesh and polyimide film, attached to a magnetic base. This sample holder was mounted to a goniometer head on macromolecular crystallography beamline, and translated along vertical and horizontal directions for raster scanning by the goniometer. Diffraction data were collected in two raster scanning approaches: (i) 100 ms X-ray exposure and 0.011° oscillation at each scan point and (ii) 500 ms X-ray exposure and 0.222° oscillation at each scan point. Using this approach, we determined the room-temperature crystal structures of lysozyme and glucose isomerase at 1.5–2.0 Å resolution. The sample holder produced negligible X-ray background scattering for data processing. Therefore, the new approach provided an opportunity to perform FT-SSX with high accessibility using macromolecular crystallography beamlines at synchrotron without any special equipment.
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Keywords: serial crystallography; fixed target; nylon mesh; enclosed film; room temperature; low-dose data collection; NAM-based sample holder serial crystallography; fixed target; nylon mesh; enclosed film; room temperature; low-dose data collection; NAM-based sample holder
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MDPI and ACS Style

Park, S.-Y.; Choi, H.; Eo, C.; Cho, Y.; Nam, K.H. Fixed-Target Serial Synchrotron Crystallography Using Nylon Mesh and Enclosed Film-Based Sample Holder. Crystals 2020, 10, 803. https://doi.org/10.3390/cryst10090803

AMA Style

Park S-Y, Choi H, Eo C, Cho Y, Nam KH. Fixed-Target Serial Synchrotron Crystallography Using Nylon Mesh and Enclosed Film-Based Sample Holder. Crystals. 2020; 10(9):803. https://doi.org/10.3390/cryst10090803

Chicago/Turabian Style

Park, Suk-Youl; Choi, Hyeongju; Eo, Cheolsoo; Cho, Yunje; Nam, Ki H. 2020. "Fixed-Target Serial Synchrotron Crystallography Using Nylon Mesh and Enclosed Film-Based Sample Holder" Crystals 10, no. 9: 803. https://doi.org/10.3390/cryst10090803

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