title: High throughput protein nanocrystal fractionation in a microfluidic sorter.
abstract: Protein crystallography is transitioning into a new generation with the introduction of the X-ray free electron laser, which can be used to solve the structures of complex proteins via serial femtosecond crystallography. Sample characteristics play a critical role in successful implementation of this new technology, whereby a small, narrow protein crystal size distribution is desired to provide high quality diffraction data. To provide such a sample, we developed a microfluidic device that facilitates dielectrophoretic sorting of heterogeneous particle mixtures into various size fractions. The first generation device demonstrated great potential and success toward this endeavor; thus, in this work, we present a comprehensive optimization study to improve throughput and control over sorting outcomes. First, device geometry was designed considering a variety of criteria, and applied potentials were modeled to determine the scheme achieving the largest sorting efficiency for isolating nanoparticles from microparticles. Further, to investigate sorting efficiency within the nanoparticle regime, critical geometrical dimensions and input parameters were optimized to achieve high sorting efficiencies. Experiments revealed fractionation of nanobeads from microbeads in the optimized device with high sorting efficiencies, and protein crystals were sorted into submicrometer size fractions as desired for future serial femtosecond crystallography experiments.
authors: Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287, United States.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, United States.
authors: Bahige G
authors: BG
authors: Abdallah
authors: Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287, United States.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, United States.
authors: Shatabdi
authors: S
authors: Roy-Chowdhury
authors: Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287, United States.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, United States.
authors: Jesse
authors: J
authors: Coe
authors: Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287, United States.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, United States.
authors: Petra
authors: P
authors: Fromme
authors: Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287, United States.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, United States.
authors: Alexandra
authors: A
authors: Ros
mesh_terms: D003461:Crystallography; D057166:High-Throughput Screening Assays; D046210:Microfluidic Analytical Techniques; D045331:Photosystem I Protein Complex / Q000737:chemistry* / Q000378:metabolism
chemical_list: D045331:Photosystem I Protein Complex