title: Differential dynamic microscopy of weakly scattering and polydisperse protein-rich clusters.
abstract: Nanoparticle dynamics impact a wide range of biological transport processes and applications in nanomedicine and natural resource engineering. Differential dynamic microscopy (DDM) was recently developed to quantify the dynamics of submicron particles in solutions from fluctuations of intensity in optical micrographs. Differential dynamic microscopy is well established for monodisperse particle populations, but has not been applied to solutions containing weakly scattering polydisperse biological nanoparticles. Here we use bright-field DDM (BDDM) to measure the dynamics of protein-rich liquid clusters, whose size ranges from tens to hundreds of nanometers and whose total volume fraction is less than 10(-5). With solutions of two proteins, hemoglobin A and lysozyme, we evaluate the cluster diffusion coefficients from the dependence of the diffusive relaxation time on the scattering wave vector. We establish that for weakly scattering populations, an optimal thickness of the sample chamber exists at which the BDDM signal is maximized at the smallest sample volume. The average cluster diffusion coefficient measured using BDDM is consistently lower than that obtained from dynamic light scattering at a scattering angle of 90°. This apparent discrepancy is due to Mie scattering from the polydisperse cluster population, in which larger clusters preferentially scatter more light in the forward direction.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas 77204-4004, USA.
authors.forename: Mohammad S
authors.initials: MS
authors.lastname: Safari
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas 77204-4004, USA.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Vorontsova
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas 77204-4004, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Poling-Skutvik
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas 77204-4004, USA.|Department of Chemistry, University of Houston, Houston, Texas 77204-4004, USA.
authors.forename: Peter G
authors.initials: PG
authors.lastname: Vekilov
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas 77204-4004, USA.
authors.forename: Jacinta C
authors.initials: JC
authors.lastname: Conrad
mesh_terms: D004058:Diffusion; D000067493:Dynamic Light Scattering; D006441:Hemoglobin A / Q000737:chemistry*; D006801:Humans; D008853:Microscopy / Q000379:methods*; D009113:Muramidase / Q000737:chemistry*; D061848:Optical Imaging / Q000379:methods; D012996:Solutions; D014783:Viscosity; D014867:Water / Q000737:chemistry
chemical_list: D012996:Solutions; D014867:Water; D006441:Hemoglobin A; D009113:Muramidase