Zhao Feng-Zhu, Sun Bo, Yu Li, Xiao Qing-Jie, Wang Zhi-Jun, Chen Liang-Liang, Liang Huan, Wang Qi-Sheng, He Jian-Hua, Yin Da-Chuan
School of Life Sciences, Northwestern Polytechnical University, Xi'an, China.
Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai, China.
Lab Chip. 2020 Oct 27;20(21):3888-3898. doi: 10.1039/d0lc00443j.
A sample delivery system is one of the key parts of serial crystallography. It is the main limiting factor affecting the application of serial crystallography. At present, although a variety of useful sample delivery systems have been developed for serial crystallography, it still remains the focus of the field to further improve the performance and efficiency of sample delivery. In existing sample delivery technologies, samples are usually delivered in linear motion. Here we show that the samples can also be delivered using circular motion, which is a novel motion mode never tested before. In this paper, we report a microfluidic rotating-target sample delivery device, which is characterized by the circular motion of the samples, and verify the performance of the device at a synchrotron radiation facility. The microfluidic rotating-target sample delivery device consists of two parts: a microfluidic sample plate and a motion control system. Sample delivery is realized by rotating the microfluidic sample plate containing in situ grown crystals. This device offers significant advantages, including a very wide adjustable range of delivery speed, low background noise, and low sample consumption. Using the microfluidic rotating-target device, we carried out in situ serial crystallography experiments with lysozyme and proteinase K as model samples at the Shanghai Synchrotron Radiation Facility, and performed structural determination based on the serial crystallographic data. The results showed that the designed device is fully compatible with the synchrotron radiation facility, and the structure determination of proteins is successful using the serial crystallographic data obtained with the device.
样品输送系统是串行晶体学的关键部件之一。它是影响串行晶体学应用的主要限制因素。目前,尽管已经为串行晶体学开发了各种有用的样品输送系统,但进一步提高样品输送的性能和效率仍然是该领域的重点。在现有的样品输送技术中,样品通常以直线运动的方式输送。在此我们表明,样品也可以使用圆周运动进行输送,这是一种以前从未测试过的新型运动模式。在本文中,我们报道了一种微流控旋转靶样品输送装置,其特点是样品做圆周运动,并在同步辐射装置上验证了该装置的性能。该微流控旋转靶样品输送装置由两部分组成:微流控样品板和运动控制系统。通过旋转包含原位生长晶体的微流控样品板来实现样品输送。该装置具有显著优势,包括输送速度的可调范围非常宽、背景噪声低以及样品消耗少。使用该微流控旋转靶装置,我们在上海同步辐射装置上以溶菌酶和蛋白酶K作为模型样品进行了原位串行晶体学实验,并基于串行晶体学数据进行了结构测定。结果表明,所设计的装置与同步辐射装置完全兼容,并且使用该装置获得的串行晶体学数据成功地完成了蛋白质的结构测定。