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Title: Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array

A microfluidic platform has been developed for the capture and X-ray analysis of protein microcrystals, affording a means to improve the efficiency of XFEL and synchrotron experiments. X-ray free-electron lasers (XFELs) promise to enable the collection of interpretable diffraction data from samples that are refractory to data collection at synchrotron sources. At present, however, more efficient sample-delivery methods that minimize the consumption of microcrystalline material are needed to allow the application of XFEL sources to a wide range of challenging structural targets of biological importance. Here, a microfluidic chip is presented in which microcrystals can be captured at fixed, addressable points in a trap array from a small volume (<10 ┬Ál) of a pre-existing slurry grown off-chip. The device can be mounted on a standard goniostat for conducting diffraction experiments at room temperature without the need for flash-cooling. Proof-of-principle tests with a model system (hen egg-white lysozyme) demonstrated the high efficiency of the microfluidic approach for crystal harvesting, permitting the collection of sufficient data from only 265 single-crystal still images to permit determination and refinement of the structure of the protein. This work shows that microfluidic capture devices can be readily used to facilitate data collection from protein microcrystals grownmore » in traditional laboratory formats, enabling analysis when cryopreservation is problematic or when only small numbers of crystals are available. Such microfluidic capture devices may also be useful for data collection at synchrotron sources.« less
Authors:
 [1] ;  [2] ;  [2] ;  [2] ;  [2] ;  [3] ;  [2] ;  [1] ;  [1] ;  [2] ; ;  [1] ;  [2] ;  [2] ;  [2] ;  [2] ; ; ;  [4] ; more »;  [5] ;  [1] ;  [2] ;  [2] ;  [2] ;  [2] ;  [6] ;  [2] « less
  1. Stanford University, Stanford, CA 94305 (United States)
  2. (United States)
  3. University of California, Berkeley, CA 94720 (United States)
  4. SLAC National Accelerator Laboratory, Stanford, CA 94305 (United States)
  5. Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States)
  6. Johns Hopkins University School of Medicine, Baltimore, MD 21205 (United States)
Publication Date:
OSTI Identifier:
22347702
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 71; Journal Issue: Pt 4; Other Information: PMCID: PMC4388268; PMID: 25849403; PUBLISHER-ID: wa5087; OAI: oai:pubmedcentral.nih.gov:4388268; Copyright (c) Lyubimov et al. 2015; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CAPTURE; COOLING; DELIVERY; EFFICIENCY; ELECTRONS; EQUIPMENT; HARVESTING; IMAGES; LYSOZYME; MONOCRYSTALS; REFRACTORIES; SYNCHROTRONS; TRAPS; X-RAY DIFFRACTION