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Simple nanofluidic devices for high-throughput, non-equilibrium studies at the single-molecule level

Carel Fijen, Mattia Fontana, Serge G. Lemay, Klaus Mathwig, Johannes Hohlbein
doi: https://doi.org/10.1101/201079
Carel Fijen
1Wageningen University and Research, Laboratory of Biophysics, Stippeneng 4, Wageningen, 6708 WE, The Netherlands
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Mattia Fontana
1Wageningen University and Research, Laboratory of Biophysics, Stippeneng 4, Wageningen, 6708 WE, The Netherlands
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Serge G. Lemay
2University of Twente, MESA+ Institute for Nanotechnology, P.O. Box 217, Enschede, 7500 AE, The Netherlands
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Klaus Mathwig
2University of Twente, MESA+ Institute for Nanotechnology, P.O. Box 217, Enschede, 7500 AE, The Netherlands
3University of Groningen, Groningen Research Institute of Pharmacy, Pharmaceutical Analysis, P.O. Box 196, 9700 AD, Groningen, The Netherlands
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  • For correspondence: k.h.mathwig@rug.nl; johannes.hohlbein@wur.nl
Johannes Hohlbein
1Wageningen University and Research, Laboratory of Biophysics, Stippeneng 4, Wageningen, 6708 WE, The Netherlands
4Wageningen University and Research, Microspectroscopy Research Facility, Stippeneng 4, Wageningen, 6708 WE, The Netherlands
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  • For correspondence: k.h.mathwig@rug.nl; johannes.hohlbein@wur.nl
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ABSTRACT

Single-molecule detection schemes offer powerful means to overcome static and dynamic heterogeneity inherent to complex samples. Probing chemical and biological interactions and reactions with high throughput and time resolution, however, remains challenging and often requires surface-immobilized entities. Here, utilizing camera-based fluorescence microscopy, we present glass-made nanofluidic devices in which fluorescently labelled molecules flow through nanochannels that confine their diffusional movement. The first design features an array of parallel nanochannels for high-throughput analysis of molecular species under equilibrium conditions allowing us to record 200.000 individual localization events in just 10 minutes. Using these localizations for single particle tracking, we were able to obtain accurate flow profiles including flow speeds and diffusion coefficients inside the channels.

A second design featuring a T-shaped nanochannel enables precise mixing of two different species as well as the continuous observation of chemical reactions. We utilized the design to visualize enzymatically driven DNA synthesis in real time and at the single-molecule level. Based on our results, we are convinced that the versatility and performance of the nanofluidic devices will enable numerous applications in the life sciences.

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Posted October 10, 2017.
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Simple nanofluidic devices for high-throughput, non-equilibrium studies at the single-molecule level
Carel Fijen, Mattia Fontana, Serge G. Lemay, Klaus Mathwig, Johannes Hohlbein
bioRxiv 201079; doi: https://doi.org/10.1101/201079
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Simple nanofluidic devices for high-throughput, non-equilibrium studies at the single-molecule level
Carel Fijen, Mattia Fontana, Serge G. Lemay, Klaus Mathwig, Johannes Hohlbein
bioRxiv 201079; doi: https://doi.org/10.1101/201079

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