In JoVE (1)
Other Publications (1)
Articles by Yannig Gicquel in JoVE
Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography Yannig Gicquel*1,2, Robin Schubert*3,4,5, Svetlana Kapis3, Gleb Bourenkov6, Thomas Schneider6, Markus Perbandt3,4, Christian Betzel3,4,5, Henry N. Chapman1,2,4, Michael Heymann1,7 1Center for Free Electron Laser Science, DESY, 2Department of Physics, University of Hamburg, 3Institute for Biochemistry and Molecular Biology, Laboratory for Structural Biology of Infection and Inflammation, University of Hamburg, 4The Hamburg Center for Ultrafast Imaging, University of Hamburg, 5Integrated Biology Infrastructure Life-Science Facility at the European XFEL (XBI), 6European Molecular Biology Laboratory, EMBL c/o DESY, 7Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry This protocol describes in detail how to fabricate and operate microfluidic devices for X-ray diffraction data collection at room temperature. Additionally, it describes how to monitor protein crystallization by dynamic light scattering and how to process and analyze obtained diffraction data.
Other articles by Yannig Gicquel on PubMed
A Multicrystal Diffraction Data-collection Approach for Studying Structural Dynamics with Millisecond Temporal Resolution IUCrJ. Nov, 2016 | Pubmed ID: 27840678 Many biochemical processes take place on timescales ranging from femto-seconds to seconds. Accordingly, any time-resolved experiment must be matched to the speed of the structural changes of interest. Therefore, the timescale of interest defines the requirements of the X-ray source, instrumentation and data-collection strategy. In this study, a minimalistic approach for crystallization is presented that requires only a few microlitres of sample solution containing a few hundred crystals. It is demonstrated that complete diffraction data sets, merged from multiple crystals, can be recorded within only a few minutes of beamtime and allow high-resolution structural information of high quality to be obtained with a temporal resolution of 40 ms. Global and site-specific radiation damage can be avoided by limiting the maximal dose per crystal to 400 kGy. Moreover, analysis of the data collected at higher doses allows the time-resolved observation of site-specific radiation damage. Therefore, our approach is well suited to observe structural changes and possibly enzymatic reactions in the low-millisecond regime.