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@ARTICLE{Gicquel:402484,
      author       = {Gicquel, Yannig and Schubert, Robin and Kapis, Svetlana and
                      Bourenkov, Gleb and Schneider, Thomas and Perbandt, Markus
                      and Betzel, Christian and Chapman, Henry N. and Heymann,
                      Michael},
      title        = {{M}icrofluidic {C}hips for {I}n {S}itu {C}rystal {X}-ray
                      {D}iffraction and {I}n {S}itu {D}ynamic {L}ight {S}cattering
                      for {S}erial {C}rystallography},
      journal      = {Journal of visualized experiments},
      volume       = {134},
      issn         = {1940-087X},
      address      = {[S.l.]},
      reportid     = {PUBDB-2018-01851},
      pages        = {e57133},
      year         = {2018},
      note         = {PIF-2015-46},
      abstract     = {This protocol describes fabricating microfluidic devices
                      with low X-ray background optimized for goniometer based
                      fixed target serial crystallography. The devices are
                      patterned from epoxy glue using soft lithography and are
                      suitable for in situ X-ray diffraction experiments at room
                      temperature. The sample wells are lidded on both sides with
                      polymeric polyimide foil windows that allow diffraction data
                      collection with low X-ray background. This fabrication
                      method is undemanding and inexpensive. After the sourcing of
                      a SU-8 master wafer, all fabrication can be completed
                      outside of a cleanroom in a typical research lab
                      environment. The chip design and fabrication protocol
                      utilize capillary valving to microfluidically split an
                      aqueous reaction into defined nanoliter sized droplets. This
                      loading mechanism avoids the sample loss from channel
                      dead-volume and can easily be performed manually without
                      using pumps or other equipment for fluid actuation. We
                      describe how isolated nanoliter sized drops of protein
                      solution can be monitored in situ by dynamic light
                      scattering to control protein crystal nucleation and
                      growth.After suitable crystals are grown, complete X-ray
                      diffraction datasets can be collected using goniometer based
                      in situ fixed target serial X-ray crystallography at room
                      temperature. The protocol provides custom scripts to process
                      diffraction datasets using a suite of software tools to
                      solve and refine the protein crystal structure. This
                      approach avoids the artefacts possibly induced during
                      cryo-preservation or manual crystal handling in conventional
                      crystallography experiments. We present and compare three
                      protein structures that were solved using small crystals
                      with dimensions of approximately 10-20 μm grown in chip. By
                      crystallizing and diffracting in situ, handling and hence
                      mechanical disturbancesof fragile crystals is minimized. The
                      protocol details how to fabricate a custom X-ray transparent
                      microfluidic chip suitable for in situ serial
                      crystallography. As almost every crystal can be used for
                      diffraction data collection, these microfluidic chips are a
                      very efficient crystal delivery method.},
      cin          = {CFEL-I / UNI/CUI / EMBL},
      ddc          = {570},
      cid          = {I:(DE-H253)CFEL-I-20161114 / $I:(DE-H253)UNI_CUI-20121230$
                      / I:(DE-H253)EMBL-20120731},
      pnm          = {6215 - Soft Matter, Health and Life Sciences (POF3-621) /
                      6G3 - PETRA III (POF3-622) / DFG project 194651731 - EXC
                      1074: Hamburger Zentrum für ultraschnelle Beobachtung
                      (CUI): Struktur, Dynamik und Kontrolle von Materie auf
                      atomarer Skala (194651731) / 05K13GU7 - Test eines "Serial
                      Femtosecond Crystallography (SFX)"-Messtandes am
                      Europäischen Elektronenlaser XFEL (BMBF-05K13GU7)},
      pid          = {G:(DE-HGF)POF3-6215 / G:(DE-HGF)POF3-6G3 /
                      G:(GEPRIS)194651731 / G:(DE-H253)BMBF-05K13GU7},
      experiment   = {EXP:(DE-H253)CFEL-Exp-20150101 /
                      EXP:(DE-H253)P-P14-20150101},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:29757285},
      UT           = {WOS:000444051300047},
      doi          = {10.3791/57133},
      url          = {https://bib-pubdb1.desy.de/record/402484},
}