title: Room-temperature serial crystallography using a kinetically optimized microfluidic device for protein crystallization and on-chip X-ray diffraction.
abstract: An emulsion-based serial crystallographic technology has been developed, in which nanolitre-sized droplets of protein solution are encapsulated in oil and stabilized by surfactant. Once the first crystal in a drop is nucleated, the small volume generates a negative feedback mechanism that lowers the supersaturation. This mechanism is exploited to produce one crystal per drop. Diffraction data are measured, one crystal at a time, from a series of room-temperature crystals stored on an X-ray semi-transparent microfluidic chip, and a 93% complete data set is obtained by merging single diffraction frames taken from different unoriented crystals. As proof of concept, the structure of glucose isomerase was solved to 2.1 Å, demonstrating the feasibility of high-throughput serial X-ray crystallography using synchrotron radiation.
authors.affiliation: Graduate Program in Biophysics and Structural Biology, Brandeis University, 415 South Street, Waltham, MA 02454, USA ; Martin Fisher School of Physics, Brandeis University, 415 South Street, Waltham, MA 02454, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Heymann
authors.affiliation: Martin Fisher School of Physics, Brandeis University, 415 South Street, Waltham, MA 02454, USA.
authors.forename: Achini
authors.initials: A
authors.lastname: Opthalage
authors.affiliation: Field of Biophysics, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Wierman
authors.affiliation: Martin Fisher School of Physics, Brandeis University, 415 South Street, Waltham, MA 02454, USA.
authors.forename: Sathish
authors.initials: S
authors.lastname: Akella
authors.affiliation: Cornell High Energy Synchrotron Source (CHESS) and Macromolecular Diffraction Facility at CHESS (MacCHESS), Cornell University, Ithaca, NY 14853, USA.
authors.forename: Doletha M E
authors.initials: DM
authors.lastname: Szebenyi
authors.affiliation: Cornell High Energy Synchrotron Source (CHESS) and Macromolecular Diffraction Facility at CHESS (MacCHESS), Cornell University, Ithaca, NY 14853, USA ; Department of Physics, Cornell University, Ithaca, NY 14853, USA ; Kavli Institute at Cornell for Nanoscale Science, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Sol M
authors.initials: SM
authors.lastname: Gruner
authors.affiliation: Martin Fisher School of Physics, Brandeis University, 415 South Street, Waltham, MA 02454, USA.
authors.forename: Seth
authors.initials: S
authors.lastname: Fraden
keywords: X-ray diffraction; microfluidic devices; protein crystallization; serial crystallography