Pearson Joseph Z, Krause Frank, Haffke Dirk, Demeler Borries, Schilling Kristian, Cölfen Helmut
Physical Chemistry, Department of Chemistry, University of Konstanz, Konstanz, Germany.
Nanolytics GmbH, Potsdam, Germany.
Methods Enzymol. 2015;562:1-26. doi: 10.1016/bs.mie.2015.06.033. Epub 2015 Aug 3.
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.
我们描述了分析超速离心（AUC）硬件方面的重要进展，这些进展为传统AUC仪器中观察到的流体动力学信息增添了新内容。与贝克曼库尔特XLA紫外/可见光检测器不同，多波长（MWL）检测不仅能够针对一个波长收集沉降数据，还能在单次实验中针对较大的波长范围收集数据。这一额外维度将数据密度提高了几个数量级，显著改善了测量的统计学结果，并为实验增添了重要信息，因为现在有了光谱表征这一额外维度来补充流体动力学信息。这种新型检测器避免了在不同波长下重复进行繁琐的实验，为基于溶液的复杂混合物研究开辟了新途径。在本章中，我们以生物聚合物为研究重点，描述了这种新型检测器设计的功能、特性及应用。我们展示了来自两种不同MWL检测器的数据，并讨论了硬件差异和不同数据采集模式的优缺点。此外，还讨论了光纤在紫外光应用中的困难。对不同平台的数据质量进行了比较。
