title: Next-Generation AUC Adds a Spectral Dimension: Development of Multiwavelength Detectors for the Analytical Ultracentrifuge.
abstract: We describe important advances in analytical ultracentrifugation (AUC) hardware, which add new information to the hydrodynamic information observed in traditional AUC instruments. In contrast to the Beckman-Coulter XLA UV/visible detector, multiwavelength (MWL) detection is able to collect sedimentation data not just for one wavelength, but for a large wavelength range in a single experiment. The additional dimension increases the data density by orders of magnitude, significantly improving the statistics of the measurement and adding important information to the experiment since an additional dimension of spectral characterization is now available to complement the hydrodynamic information. The new detector avoids tedious repeats of experiments at different wavelengths and opens up new avenues for the solution-based investigation of complex mixtures. In this chapter, we describe the capabilities, characteristics, and applications of the new detector design with biopolymers as the focus of study. We show data from two different MWL detectors and discuss strengths and weaknesses of differences in the hardware and different data acquisition modes. Also, difficulties with fiber optic applications in the UV are discussed. Data quality is compared across platforms.
authors.affiliation: Physical Chemistry, Department of Chemistry, University of Konstanz, Konstanz, Germany.
authors.forename: Joseph Z
authors.initials: JZ
authors.lastname: Pearson
authors.affiliation: Nanolytics GmbH, Potsdam, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Krause
authors.affiliation: Physical Chemistry, Department of Chemistry, University of Konstanz, Konstanz, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Haffke
authors.affiliation: Department of Biochemistry, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA.
authors.forename: Borries
authors.initials: B
authors.lastname: Demeler
authors.affiliation: Nanolytics GmbH, Potsdam, Germany.
authors.forename: Kristian
authors.initials: K
authors.lastname: Schilling
authors.affiliation: Physical Chemistry, Department of Chemistry, University of Konstanz, Konstanz, Germany. Electronic address: Helmut.Coelfen@uni-konstanz.de.
authors.forename: Helmut
authors.initials: H
authors.lastname: Cölfen
mesh_terms: D000818:Animals; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D006454:Hemoglobins / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification*; D059629:Signal-To-Noise Ratio; D013056:Spectrophotometry, Ultraviolet / Q000295:instrumentation / Q000379:methods; D014461:Ultracentrifugation / Q000295:instrumentation / Q000379:methods
chemical_list: D006454:Hemoglobins; D012710:Serum Albumin, Bovine; D004247:DNA
keywords: Analytical ultracentrifugation; Hydrodynamic analysis; Multiwavelength detector; Optical detector development; Spectral analysis; UV–vis absorbance