Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography

作者:Roessler Christian G; Agarwal Rakhi; Allaire Marc; Alonso Mori Roberto; Andi Babak; Bachega Jose F R; Bommer Martin; Brewster Aaron S; Browne Michael C; Chatterjee Ruchira; Cho Eunsun; Cohen Aina E; Cowan Matthew; Datwani Sammy; Davidson Victor L; Defever Jim; Eaton Brent; Ellson Richard; Feng Yiping; Ghislain Lucien P; Glownia James M; Han Guangye; Hattne Johan; Hellmich Julia; Heroux Annie; Ibrahim Mohamed; Kern Jan; Kuczewski Anthony; Lemke Henrik T
来源:Structure, 2016, 24(4): 631-640.
DOI:10.1016/j.str.2016.02.007

摘要

X-ray free-electron lasers (XFELs) provide very intense X-ray pulses suitable for macromolecular crystallography. Each X-ray pulse typically lasts for tens of femtoseconds and the interval between pulses is many orders of magnitude longer. Here we describe two novel acoustic injection systems that use focused sound waves to eject picoliter to nanoliter crystal-containing droplets out of microplates and into the X-ray pulse from which diffraction data are collected. The on-demand droplet delivery is synchronized to the XFEL pulse scheme, resulting in X-ray pulses intersecting up to 88% of the droplets. We tested several types of samples in a range of crystallization conditions, wherein the overall crystal hit ratio (e.g., fraction of images with observable diffraction patterns) is a function of the microcrystal slurry concentration. We report crystal structures from lysozyme, thermolysin, and stachydrine demethylase (Stc2). Additional samples were screened to demonstrate that these methods can be applied to rare samples.

  • 出版日期2016-4-5