4.2 Article

The HARE chip for efficient time-resolved serial synchrotron crystallography

Journal

JOURNAL OF SYNCHROTRON RADIATION
Volume 27, Issue -, Pages 360-370

Publisher

INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S1600577520000685

Keywords

data collection; sample delivery; fixed-target serial synchrotron crystallography; time-resolved crystallography; HARE; LAMA

Funding

  1. Max Planck Society
  2. Cluster of Excellence 'The Hamburg Centre for Ultrafast Imaging' of the Deutsche Forschungsgemeinschaft [EXC 1074, 194651731]
  3. European Union [623994]
  4. Joachim Herz Foundation (Biomedical Physics of Infection)
  5. Alexander von Humboldt-Stiftung postdoctoral research award
  6. BMBF ('Verbundforschung') [05K16GU1, 05K19GU1]

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Serial synchrotron crystallography (SSX) is an emerging technique for static and time-resolved protein structure determination. Using specifically patterned silicon chips for sample delivery, the 'hit-and-return' (HARE) protocol allows for efficient time-resolved data collection. The specific pattern of the crystal wells in the HARE chip provides direct access to many discrete time points. HARE chips allow for optical excitation as well as on-chip mixing for reaction initiation, making a large number of protein systems amenable to time-resolved studies. Loading of protein microcrystals onto the HARE chip is streamlined by a novel vacuum loading platform that allows fine-tuning of suction strength while maintaining a humid environment to prevent crystal dehydration. To enable the widespread use of time-resolved serial synchrotron crystallography (TR-SSX), detailed technical descriptions of a set of accessories that facilitate TR-SSX workflows are provided.

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