High-throughput counter-diffusion capillary crystallization and in situ diffraction using high-pressure freezing in protein crystallography

• Published in: Journal of applied crystallography Vol. 45; no. 5; pp. 999 - 1008
• Main Authors: Kurz, Mareike, Blattmann, Beat, Kaech, Andres, Briand, Christophe, Reardon, Paul, Ziegler, Urs, Gruetter, Markus G.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01-10-2012; Blackwell Publishing Ltd
• Subjects: Capillarity; Collection; counter-diffusion capillary crystallization; Cryoprotectants; Crystallization; Crystallography; Crystals; Diffraction; Freezing; high-pressure freezing; high-throughput techniques; Molecular biology; Nanostructure; protein crystallography; Proteins
• ISSN: 1600-5767; 0021-8898; 1600-5767
• DOI: 10.1107/S0021889812034061

Post‐crystallization treatments such as manual fishing of crystals and soaking in cryoprotectant solutions, especially of large macromolecular complexes and membrane proteins, are cumbersome and often lead to crystal damage and reduced diffraction data quality. Here, a capillary crystallization plate is presented that simultaneously allows counter‐diffusion crystallization at the nanolitre scale in a high‐throughput screening mode, low‐temperature in situ diffraction data collection from crystals after cryoprotection and low‐temperature in situ data collection of crystals without the addition of any cryoprotectant after high‐pressure (HP) freezing. The development of this plate and plunge cooling of crystals in the capillaries is a major step towards implementing automated in situ high‐throughput crystal diffraction data collection at a synchrotron beamline. In combination with HP freezing this offers a new opportunity to obtain structural information from fragile crystals of supramolecular complexes that might otherwise not be feasible.