Out-of-equilibrium biomolecular interactions monitored by picosecond fluorescence in microfluidic droplets

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-equilib...

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Bibliographic Details
Published in:Lab on a chip 2014-01, Vol.14 (10), p.1767-1774
Main Authors: Maillot, Sacha, Carvalho, Alain, Vola, Jean-Pierre, Boudier, Christian, Mély, Yves, Haacke, Stefan, Léonard, Jérémie
Format: Article
Language:English
Subjects:
Animals
Cattle
Condensed Matter
Fluorescent Dyes - chemistry
Kinetics
Microfluidic Analytical Techniques - instrumentation
Microfluidic Analytical Techniques - methods
Oils - chemistry
Physics
Rosaniline Dyes - chemistry
Serum Albumin, Bovine - chemistry
Serum Albumin, Bovine - metabolism
Signal-To-Noise Ratio
Spectrometry, Fluorescence
Time Factors
Water - chemistry
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Summary: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.
ISSN:1473-0197
1473-0189
1473-0189
DOI:10.1039/c3lc51283e