High Throughput Protein Nanocrystal Fractionation in a Microfluidic Sorter
mediaposted on 2015-04-21, 00:00 authored by Bahige G. Abdallah, Shatabdi Roy-Chowdhury, Jesse Coe, Petra Fromme, Alexandra Ros
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.
Microfluidic SorterProtein crystallographysize fractionsquality diffraction dataelectron laserHigh Throughput Protein Nanocrystal Fractionationmicrofluidic deviceprotein crystalsoptimized deviceSample characteristicssubmicrometer size fractionsfemtosecond crystallography experimentsparticle mixturesdevice geometryefficiencyfemtosecond crystallographyoptimization studygeneration devicenanoparticle regimeprotein crystal size distributioninput parameters