同步辐射串行晶体学静电纺丝上样实验技术

串行晶体学作为一种解析蛋白质晶体结构的新方法,因其拥有室温采集、辐射损伤低、时间分辨等优势而得到迅速发展。早期发展于自由电子激光的串行飞秒晶体学已成功实现了蛋白质微小晶体结构的解析。为了引入串行晶体学的技术优势,充分发挥第三代同步辐射的丰富资源和特点,基于上海光源BL17U1生物大分子线站,我们对串行晶体学上样方法进行了理论和实验研究,解决了静电纺丝上样方式在同步辐射上应用的难题,用溶菌酶微小晶体论证了其可行性;同时探讨了基于同步辐射的串行晶体学在毫秒时间尺度上的发展潜力,为光束线技术升级提供参考。

Electrospinning sample delivery for serial crystallography using synchrotron radiation

Background: As a novel approach to solve the protein structure, serial femtosecond crystallography (SFX) has been developed rapidly owing to the advantages of the room-temperature data collection, lower radiation damage and time-resolved study. The first X-ray free-electron laser (XFEL) based SFX experiment has offered a new method to structure determination of protein microcrystals successfully. Purpose: The aim is to introduce SFX into the widely-available third-generation synchrotron radiation (SR). Methods: The electrospinning sample delivery system is a potential method to be applied on SR. The theoretical and experimental researches on electrospinning were studied using BL17U1 beamline in Shanghai synchrotron radiation facility (SSRF). Results: A microjet containing microcrystals was produced from the liquid meniscus and controlled by an electrostatic field. The air ionization problem disturbing electrospinning in SR experiment was solved by providing vacuum condition and coaxial electrospinning technique. Using lysozyme microcrystals as a model sample, the feasibility of electrospinning was demonstrated. Conclusion: It is shown that the serial crystallography on millisecond time scale is suitable for SR, which provides a reference for the future beamline upgrading.