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Issue 1, 2019
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High-throughput, non-equilibrium studies of single biomolecules using glass-made nanofluidic devices

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Abstract

Single-molecule detection schemes offer powerful means to overcome static and dynamic heterogeneity inherent to complex samples. However, probing biomolecular interactions and reactions with high throughput and time resolution remains challenging, often requiring surface-immobilized entities. Here, we introduce glass-made nanofluidic devices for the high-throughput detection of freely-diffusing single biomolecules by camera-based fluorescence microscopy. Nanochannels of 200 nm height and a width of several micrometers confine the movement of biomolecules. Using pressure-driven flow through an array of parallel nanochannels and by tracking the movement of fluorescently labelled DNA oligonucleotides, we observe conformational changes with high throughput. In a device geometry featuring a T-shaped junction of nanochannels, we drive steady-state non-equilibrium conditions by continuously mixing reactants and triggering chemical reactions. We use the device to probe the conformational equilibrium of a DNA hairpin as well as to continuously observe DNA synthesis in real time. Our platform offers a straightforward and robust method for studying reaction kinetics at the single-molecule level.

Graphical abstract: High-throughput, non-equilibrium studies of single biomolecules using glass-made nanofluidic devices

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Supplementary files

Article information


Submitted
29 Oct 2018
Accepted
20 Nov 2018
First published
20 Nov 2018

Lab Chip, 2019,19, 79-86
Article type
Paper

High-throughput, non-equilibrium studies of single biomolecules using glass-made nanofluidic devices

M. Fontana, C. Fijen, S. G. Lemay, K. Mathwig and J. Hohlbein, Lab Chip, 2019, 19, 79
DOI: 10.1039/C8LC01175C

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