Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography

Journal Article · Fri Apr 01 00:00:00 EDT 2016 · Structure

DOI:https://doi.org/10.1016/j.str.2016.02.007· OSTI ID:1349763

Roessler, Christian G.; Agarwal, Rakhi; Allaire, Marc; Alonso-Mori, Roberto; Andi, Babak; Bachega, José 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; Héroux, Annie; Ibrahim, Mohamed; Kern, Jan; Kuczewski, Anthony; Lemke, Henrik T.; Liu, Pinghua; Majlof, Lars; McClintock, William M.; Myers, Stuart; Nelsen, Silke; Olechno, Joe; Orville, Allen M.; Sauter, Nicholas K.; Soares, Alexei S.; Soltis, S. Michael; Song, Heng; Stearns, Richard G.; Tran, Rosalie; Tsai, Yingssu; Uervirojnangkoorn, Monarin; Wilmot, Carrie M.; Yachandra, Vittal; Yano, Junko; Yukl, Erik T.; Zhu, Diling; Zouni, Athina

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.

• Research Organization: Diamond Light Source Ltd., Oxfordshire (United Kingdom); Brookhaven National Laboratory (BNL), Upton, NY (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
• Sponsoring Organization: USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Biological and Environmental Research (BER)
• Grant/Contract Number: AC02-76SF00515; 11-008; FWP BO-70; BO-9734; AC02-05CH11231; AC02-98CH10886; SC00112704
• OSTI ID: 1349763
• Alternate ID(s): OSTI ID: 1253633; OSTI ID: 1337424; OSTI ID: 1354254; OSTI ID: 1379277
• Report Number(s): BNL-112770-2016-JA; S0969212616000526; PII: S0969212616000526
• Journal Information: Structure, Journal Name: Structure Vol. 24 Journal Issue: 4; ISSN 0969-2126
• Publisher: Elsevier
• Country of Publication: United Kingdom
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY