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

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 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 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 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.

Topics

• density
• polymer
• phase
• experiment
• crystallization