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2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Growing Protein Crystals with Distinct Dimensions Using Automated Crystallization Coupled with In Situ Dynamic Light Scattering

Daniela Baitan^1,2, Robin Schubert^2,3, Arne Meyer¹, Karsten Dierks¹, Markus Perbandt^2,3, Christian Betzel^2,3

¹Xtal Concepts GmbH, ²Institute for Biochemistry and Molecular Biology, Laboratory for Structural Biology of Infection and Inflammation c/o DESY, University of Hamburg, ³The Hamburg Center for Ultrafast Imaging, University of Hamburg

Here we present a protocol for controlled production of protein microcrystals. The process uses an automated device allowing controlled manipulation of several crystallization parameters. The protein crystallization is carried-out by controlled and automated addition of crystallization solutions while monitoring and investigating the radius distribution of particles in the crystallization droplet.

Chemistry

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 Perbandt^3,4, Christian Betzel^3,4,5, Henry N. Chapman^1,2,4, Michael Heymann^1,7

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.