High-density grids for efficient data collection from multiple crystals

Elizabeth L. Baxter, Laura Aguila, Roberto Alonso-Mori, Christopher O. Barnes, Christopher A. Bonagura, Winnie Brehmer, Axel T. Brunger, Guillermo Calero, Tom T. Caradoc-Davies, Ruchira Chatterjee, William F. Degrado, James S. Fraser, Mohamed Ibrahim, Jan Kern, Brian K. Kobilka, Andrew C. Kruse, Karl M. Larsson, Heinrik T. Lemke, Artem Y. Lyubimov, Aashish ManglikScott E. McPhillips, Erik Norgren, Siew S. Pang, S. M. Soltis, Jinhu Song, Jessica Thomaston, Yingssu Tsai, William I. Weis, Rahel A. Woldeyes, Vittal Yachandra, Junko Yano, Athina Zouni, Aina E. Cohen

Research output: Contribution to journalArticleResearchpeer-review

43 Citations (Scopus)

Abstract

Higher throughput methods to mount and collect data from multiple small and radiation-sensitive crystals are important to support challenging structural investigations using microfocus synchrotron beamlines. Furthermore, efficient sample-delivery methods are essential to carry out productive femtosecond crystallography experiments at X-ray free-electron laser (XFEL) sources such as the Linac Coherent Light Source (LCLS). To address these needs, a highdensity sample grid useful as a scaffold for both crystal growth and diffraction data collection has been developed and utilized for efficient goniometer-based sample delivery at synchrotron and XFEL sources. A single grid contains 75 mounting ports and fits inside an SSRL cassette or uni-puck storage container. The use of grids with an SSRL cassette expands the cassette capacity up to 7200 samples. Grids may also be covered with a polymer film or sleeve for efficient room-temperature data collection from multiple samples. New automated routines have been incorporated into the Blu-Ice/DCSS experimental control system to support grids, including semi-automated grid alignment, fully automated positioning of grid ports, rastering and automated data collection. Specialized tools have been developed to support crystallization experiments on grids, including a universal adaptor, which allows grids to be filled by commercial liquid-handling robots, as well as incubation chambers, which support vapordiffusion and lipidic cubic phase crystallization experiments. Experiments in which crystals were loaded into grids or grown on grids using liquid-handling robots and incubation chambers are described. Crystals were screened at LCLSXPP and SSRL BL12-2 at room temperature and cryogenic temperatures.

Original languageEnglish
Pages (from-to)2-11
Number of pages10
JournalActa Crystallographica Section D: Structural Biology
VolumeD72
DOIs
Publication statusPublished - 2016

Keywords

  • Automation for sample-exchange robots
  • High-throughput crystallography
  • Sample delivery
  • Serial crystallography
  • XFELs

Cite this