Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography.

Abstract

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.

DOI: 10.1016/j.str.2016.02.007

Cite this paper

@article{Roessler2016AcousticIF, title={Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography.}, author={Christian G Roessler and Rakhi Agarwal and Marc Allaire and Roberto Alonso-Mori and Babak Andi and Jos{\'e} F R Bachega and Martin Bommer and Aaron S Brewster and Michael C Browne and Ruchira Chatterjee and Eunsun Cho and Aina E Cohen and Matthew Cowan and Sammy Datwani and Victor L Davidson and Jim Defever and Brent Eaton and Richard Ellson and Yiping Feng and Lucien P Ghislain and James M Glownia and Guangye Han and Johan Hattne and Julia Hellmich and Annie H{\'e}roux and Mohamed Ibrahim and Jan Kern and Anthony Kuczewski and Henrik T Lemke and Pinghua Liu and Lars Majlof and William M McClintock and Stuart Myers and Silke Nelsen and Joe Olechno and Allen M Orville and Nicholas K Sauter and Alexei S Soares and S Michael Soltis and Heng Song and Richard G Stearns and Rosalie Tran and Yingssu Tsai and Monarin Uervirojnangkoorn and Carrie M Wilmot and Vittal Yachandra and Junko Yano and Erik T Yukl and Diling Zhu and Athina Zouni}, journal={Structure}, year={2016}, volume={24 4}, pages={631-640} }