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

Mattia Fontana, Carel Fijen, Serge G. Lemay, Klaus Mathwig*, Johannes Hohlbein

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)
1 Downloads (Pure)

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.

Original languageEnglish
Pages (from-to)79-86
Number of pages8
JournalLab on a chip
Volume19
Issue number1
DOIs
Publication statusPublished - 7 Jan 2019

Fingerprint

Nanofluidics
Biomolecules
Glass
DNA
Throughput
Equipment and Supplies
Molecules
Oligonucleotides
Fluorescence microscopy
Fluorescence Microscopy
Reaction kinetics
Chemical reactions
Cameras
Pressure
Geometry

Cite this

Fontana, Mattia ; Fijen, Carel ; Lemay, Serge G. ; Mathwig, Klaus ; Hohlbein, Johannes. / High-throughput, non-equilibrium studies of single biomolecules using glass-made nanofluidic devices. In: Lab on a chip. 2019 ; Vol. 19, No. 1. pp. 79-86.
@article{6751561fbf4249df8a06b669d78c8b50,
title = "High-throughput, non-equilibrium studies of single biomolecules using glass-made nanofluidic devices",
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.",
author = "Mattia Fontana and Carel Fijen and Lemay, {Serge G.} and Klaus Mathwig and Johannes Hohlbein",
year = "2019",
month = "1",
day = "7",
doi = "10.1039/c8lc01175c",
language = "English",
volume = "19",
pages = "79--86",
journal = "Lab on a chip",
issn = "1473-0197",
publisher = "Royal Society of Chemistry",
number = "1",

}

High-throughput, non-equilibrium studies of single biomolecules using glass-made nanofluidic devices. / Fontana, Mattia; Fijen, Carel; Lemay, Serge G.; Mathwig, Klaus; Hohlbein, Johannes.

In: Lab on a chip, Vol. 19, No. 1, 07.01.2019, p. 79-86.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - High-throughput, non-equilibrium studies of single biomolecules using glass-made nanofluidic devices

AU - Fontana, Mattia

AU - Fijen, Carel

AU - Lemay, Serge G.

AU - Mathwig, Klaus

AU - Hohlbein, Johannes

PY - 2019/1/7

Y1 - 2019/1/7

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85058764977&partnerID=8YFLogxK

U2 - 10.1039/c8lc01175c

DO - 10.1039/c8lc01175c

M3 - Article

C2 - 30468446

AN - SCOPUS:85058764977

VL - 19

SP - 79

EP - 86

JO - Lab on a chip

JF - Lab on a chip

SN - 1473-0197

IS - 1

ER -