High-density grids for efficient data collection from multiple crystals

作者：Baxter Elizabeth L; Aguila Laura; Alonso Mori Roberto; Barnes Christopher O; Bonagura Christopher A; Brehmer Winnie; Brunger Axel T; Calero Guillermo; Caradoc Davies Tom T; Chatterjee Ruchira; Degrado William F; Fraser James S; Ibrahim Mohamed; Kern Jan; Kobilka Brian K; Kruse Andrew C; Larsson Karl M; Lemke Heinrik T; Lyubimov Artem Y; Manglik Aashish; McPhillips Scott E; Norgren Erik; Pang Siew S; Soltis S M; Song Jinhu; Thomaston Jessica

来源：Acta Crystallographica Section D-Structural Biology, 2016, 72(1): 2-11.

DOI：10.1107/S2059798315020847

• XFELs
• high-throughput crystallography
• serial crystallography
• sample delivery
• automation for sample-exchange robots

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 high-density 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 theBlu-Ice Blu-Ice/DCSS 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 vapor-diffusion 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 LCLS-XPP and SSRL BL12-2 at room temperature and cryogenic temperatures.

• 出版日期2016-1