Out-of-equilibrium biomolecular interactions monitored by picosecond fluorescence in microfluidic droplets.
Fiche publication
Date publication
mai 2014
Auteurs
Membres identifiés du Cancéropôle Est :
Pr MELY Yves
Tous les auteurs :
Maillot S, Carvalho A, Vola JP, Boudier C, Mely Y, Haacke S, Leonard J
Lien Pubmed
Résumé
We developed a new experimental approach combining Time-Resolved Fluorescence (TRF) spectroscopy and Droplet Microfluidics (DmuF) to investigate the relaxation dynamics of structurally heterogeneous biomolecular systems. Here DmuF 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.
Référence
Lab Chip. 2014 May 21;14(10):1767-74