Institut de Physique et Chimie des Matériaux de Strasbourg & Labex NIE, Université de Strasbourg, CNRS UMR 7504, F-67034 Strasbourg Cedex 2, France.
Lab Chip. 2014 May 21;14(10):1767-74. doi: 10.1039/c3lc51283e. Epub 2014 Mar 28.
We developed a new experimental approach combining Time-Resolved Fluorescence (TRF) spectroscopy and Droplet Microfluidics (DμF) to investigate the relaxation dynamics of structurally heterogeneous biomolecular systems. Here DμF was used to produce with minimal material consumption an out-of-equilibrium, fluorescently labeled biomolecular complex by rapid mixing within the droplets. TRF detection was implemented with a streak camera to monitor the time evolution of the structural heterogeneity of the complex along its relaxation towards equilibrium while it propagates inside the microfluidic channel. The approach was validated by investigating the fluorescence decay kinetics of a model interacting system of bovine serum albumin and Patent Blue V. Fluorescence decay kinetics are acquired with very good signal-to-noise ratio and allow for global, multicomponent fluorescence decay analysis, evidencing heterogeneous structural relaxation over several 100 ms.
我们开发了一种新的实验方法，结合时间分辨荧光（TRF）光谱和液滴微流控（DμF）来研究结构异质的生物分子系统的弛豫动力学。在这里，DμF 用于通过在液滴内快速混合以最小的材料消耗来产生非平衡的、荧光标记的生物分子复合物。TRF 检测采用条纹相机来监测复合物在沿着其弛豫到平衡的过程中沿着微流道传播时的结构异质性的时间演变。该方法通过研究牛血清白蛋白和专利蓝 V 的模型相互作用系统的荧光衰减动力学来验证。荧光衰减动力学具有非常好的信噪比，并允许进行全局、多组分荧光衰减分析，证明了在几百毫秒的时间内存在异质结构弛豫。
