NANOFLUIDIC-SMFD

Nanofluidic devices for high-throughput single-molecule-fluorescence detection

Coordinatore	WAGENINGEN UNIVERSITY    more  Organization address address: DROEVENDAALSESTEEG 4 city: WAGENINGEN postcode: 6708 PB contact info Titolo: Mr. Nome: Tom Cognome: Bessems Email: send email Telefono: +31 317 484478
Nazionalità Coordinatore	Netherlands [NL]
Totale costo	100˙000 €
EC contributo	100˙000 €
Programma	FP7-PEOPLE Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013)
Code Call	FP7-PEOPLE-2013-CIG
Funding Scheme	MC-CIG
Anno di inizio	2014
Periodo (anno-mese-giorno)	2014-03-01   -   2018-02-28

 Partecipanti

#	participant	country	role	EC contrib. [€]
1	WAGENINGEN UNIVERSITY  Organization address address: DROEVENDAALSESTEEG 4 city: WAGENINGEN postcode: 6708 PB contact info Titolo: Mr. Nome: Tom Cognome: Bessems Email: send email Telefono: +31 317 484478	NL (WAGENINGEN)	coordinator	100˙000.00

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 Obiettivo del progetto (Objective)

'Currently, the two prominent schemes for single-molecule fluorescence detection (SMFD), confocal microscopy and camera-based total-internal-reflection or wide-field microscopy, are ultimately limited in their ability to combine the detection of many molecules with obtaining data at sufficiently high time resolution, necessary for resolving fast dynamics with single-molecule fluorescence resonance energy transfer (smFRET). In particular, monitoring enzymatic reactions using smFRET is extremely challenging and remains to a large extent unexplored.

Here, I propose a novel nanofluidic device to overcome these limitations by using nanochannels, which provide a well-defined flow path for a fluorescent species through the excitation/detection focus of a conventional wide-field microscope. Using an array of nanochannels offers several advantages: First, the geometrical confinement enables long observation times of non-immobilized molecules. Second, the residence time of molecules in the channels is easily controlled by the flow velocity set by the syringe pump. Third, faster flow rates together with using a CCD camera in ‘streaking mode’ enable a sub-millisecond time resolution. Fourth, a high-throughput detection is achieved by using a parallel array of channels. Fifth, enzymatic reactions can be directly triggered by mixing necessary components on-the-fly using an additional inlet.

The described device will pave the way for high-throughput single-molecule detection, which will greatly expand the possibilities for researchers to apply single-molecule methods in the area of Life Sciences.'