Lautenbach Vanessa, Onishchukov Georgy, Wawra Simon E, Frank Uwe, Hartmann Lukas, Peukert Wolfgang, Walter Johannes
Institute of Particle Technology (LFG), Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU) Cauerstraße 4 91058 Erlangen Germany
Interdisciplinary Center for Functional Particle Systems (FPS), Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU) Haberstraße 9a 91058 Erlangen Germany.
Nanoscale Adv. 2024 Feb 19;6(10):2611-2622. doi: 10.1039/d3na00980g. eCollection 2024 May 14.
An advanced design of the analytical ultracentrifuge with multiwavelength emission detection (MWE-AUC) is presented which offers outstanding performance concerning the spectral resolution and range flexibility as well as the quality of the data acquired. The excitation by a 520 nm laser is complemented with a 405 nm laser. An external spectrograph with three switchable tunable gratings permits optimisation of the spectral resolution in an order of magnitude range while keeping the spectral region broad. The new system design leads also to a significant reduction of systematic signal noise and allows the assessment and control of inner filter effects. Details regarding the very large signal dynamic range are presented, an important aspect when studying samples in a broad concentration range of up to five orders of magnitude. Our system is validated by complementary studies on two biological systems, fluorescent BSA and GFP, using the commercial Optima AUC with absorbance detection for comparison. Finally, we demonstrate the capabilities of our second generation MWE-AUC with respect to multiwavelength characterisation of gold nanoclusters, which exhibit specific fluorescence depending on their structure. Overall, this work depicts an important stepping stone for the concept of multiwavelength emission detection in AUC. The MWE-AUC developed, being to our knowledge the first and sole one of its kind, has reached the development level suitable for the future in-depth studies of size-, shape- and composition-dependent emission properties of colloids.
本文介绍了一种采用多波长发射检测技术的先进分析型超速离心机（MWE-AUC），该离心机在光谱分辨率、范围灵活性以及所采集数据的质量方面表现出色。520 nm激光激发辅以405 nm激光。带有三个可切换可调光栅的外部光谱仪可在保持光谱区域宽广的同时，将光谱分辨率优化一个数量级范围。新的系统设计还显著降低了系统信号噪声，并允许对内滤效应进行评估和控制。文中介绍了非常大的信号动态范围的细节，这在研究浓度范围高达五个数量级的样品时是一个重要方面。我们的系统通过对两种生物系统（荧光牛血清白蛋白和绿色荧光蛋白）的补充研究进行了验证，并与使用吸光度检测的商用Optima AUC进行了比较。最后，我们展示了第二代MWE-AUC在金纳米团簇多波长表征方面的能力，金纳米团簇根据其结构表现出特定的荧光。总体而言，这项工作描绘了AUC中多波长发射检测概念的一个重要基石。据我们所知，所开发的MWE-AUC是同类中的首个也是唯一的一个，已达到适合未来深入研究胶体尺寸、形状和组成依赖性发射特性的发展水平。
