期刊
JOURNAL OF APPLIED CRYSTALLOGRAPHY
卷 54, 期 -, 页码 1081-1087出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S1600576721005720
关键词
serial crystallography; sample delivery; viscous media; microfluidics; polyimides
资金
- National Research Foundation of Korea [NRF-2017M3A9F6029736]
- National Research Foundation of Korea [PAL-2021] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Serial crystallography (SX) allows for room-temperature crystal structures with minimal radiation damage and molecular dynamics comprehension. A stable system for sample delivery in a viscous medium via a polyimide-based single-channel microfluidic (PSM) chip was developed, successfully determining room-temperature lysozyme crystal structures at 1.85 angstrom resolution. Ultimately, this method greatly facilitates sample delivery for SX experiments using synchrotron X-rays.
Serial crystallography (SX) provides room-temperature crystal structures with minimal radiation damage and facilitates the comprehension of molecular dynamics through time-resolved studies. In SX experiments, it is important to deliver a large number of crystal samples to the X-ray interaction point in a serial and stable manner. The advantage of crystal delivery in a viscous medium via a capillary is the ability to deliver all of the crystal samples to the X-ray interaction point at a low flow rate; however, the capillary often breaks during handling and high X-ray absorption can occur at low energy states. This study aimed to develop a stable system for sample delivery in a viscous medium via a polyimide-based single-channel microfluidic (PSM) chip for SX. Since this microfluidic chip comprises a polyimide film, it has high tensile strength and higher X-ray transmittance than a quartz capillary. The PSM chip was connected to a syringe containing the microcrystals embedded in viscous medium. The channel of the PSM chip was aligned to the X-ray path, and the viscous medium containing lysozyme crystals was stably delivered using a syringe pump at a flow rate of 100 nl min(-1). Room-temperature lysozyme crystal structures were successfully determined at 1.85 angstrom resolution. This method would greatly facilitate sample delivery for SX experiments using synchrotron X-rays.
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