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使用粘性介质、注射器和注射器泵进行连续结晶学的样品输送。

Sample delivery using viscous media, a syringe and a syringe pump for serial crystallography.

机构信息

Pohang Accelerator Laboratory, POSTECH, Pohang, Gyeongbuk 37673, Republic of Korea.

Division of Biotechnology, Korea University, Seoul 02841, Republic of Korea.

出版信息

J Synchrotron Radiat. 2019 Sep 1;26(Pt 5):1815-1819. doi: 10.1107/S160057751900897X. Epub 2019 Aug 15.

Abstract

Sample delivery using injectors is widely used in serial crystallography (SX) and has significantly contributed to the determination of crystal structures at room temperature. However, sophisticated injector nozzle fabrication methods and sample delivery operations have made it difficult for ordinary users to access the SX research. Herein, a simple and easily accessible sample delivery method for SX experiments is introduced, that uses a viscous medium, commercially available syringe and syringe pump. The syringe containing the lysozyme crystals embedded in lipidic cubic phase (LCP) or polyacrylamide (PAM) delivery media was connected to a needle having an inner diameter of 168 µm, after which it was installed on a syringe pump. By driving the syringe pump, the syringe plunger was pushed and the crystal sample was delivered to the X-ray beam position in a stable manner. Using this system, the room-temperature crystal structures of lysozyme embedded in LCP and PAM at 1.56 Å and 1.75 Å, respectively, were determined. This straightforward syringe pump-based sample delivery system can be utilized in SX.

摘要

采用注射器进样的方法在序列晶体学(SX)中得到了广泛的应用,并且为在室温下确定晶体结构做出了重要贡献。然而,复杂的注射器喷嘴制造方法和进样操作使得普通用户难以接触到 SX 研究。在此,我们介绍了一种简单易用的 SX 实验进样方法,该方法使用粘性介质、市售注射器和注射器泵。将含有嵌入在类脂立方相(LCP)或聚丙烯酰胺(PAM)中的溶菌酶晶体的注射器与内径为 168 µm 的针头连接,然后将其安装在注射器泵上。通过驱动注射器泵,推动注射器柱塞,以稳定的方式将晶体样品输送到 X 射线光束位置。使用该系统,分别在室温下确定了嵌入在 LCP 和 PAM 中的溶菌酶的晶体结构,分辨率分别为 1.56 Å 和 1.75 Å。这种基于简单注射器泵的样品输送系统可用于 SX。

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