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Rapid Nanofluidics Valves for Single-Molecule Imaging

Descrizione del progetto

Una valvola nanofluidica rapida consente l’imaging delle interazioni fra singole molecole in vivo

L’imaging delle attività di singole molecole richiede attualmente modifiche chimiche o l’immobilizzazione della superficie per agevolare la visualizzazione. Tuttavia, le aberrazioni biochimiche e cinetiche ostacolano la comprensione delle funzioni e dei processi in vivo. Grazie al sostegno del programma di azioni Marie Skłodowska-Curie, il progetto RAVASI migliorerà le capacità dell’innovativo sistema di microscopia ottica a diffusione nanofluidica sviluppato con l’obiettivo di agevolare il processo di imaging. Attualmente le molecole non trascorrono un tempo sufficiente nel canale nanofluidico per poter gestire un numero statisticamente rilevante di interazioni. Una valvola nanofluidica rapida consentirà di controllare il confinamento e il rilascio delle molecole rilevanti per superare questo ostacolo.

Obiettivo

A major challenge in biomolecular research is the investigation of biomolecular interaction at single-molecule level. Biomolecules possess heterogeneities of high physiological relevance that can only be unravelled with single-molecule tools. All current single-molecule imaging methods, however, require chemical modifications, such as fluorescent labelling or immobilization onto a surface, which might alter the biomolecule’s natural behaviour.

Recently, I have developed a ground-breaking optical microscopy method – Nanofluidic Scattering Microscopy (NSM) – whose unprecedented resolution enabled me to bypass those limitations and to image individual small proteins in free motion without any label. Despite these attractive attributes, in its current form, NSM does not allow for quantitative study of interaction kinetics between individual molecules. Due to the inherently fast Brownian motion, the time that a molecule spends on average in the optically probed volume – a nanofluidic channel – is substantially shorter than the time required to record a statistically relevant number of association and dissociation events.

In this project, we will develop an essential nanoscopic component – a rapid nanofluidic valve – that will enable to confine and release interacting biomolecules to and from the nanofluidic volume. The nanofluidic valves will be based on the principles of thermo-responsive polymer hydrogel in combination with nanoplasmonic heating. The integration of the nanofluidic valves with NSM will enable to track evolution of individual biomolecules at single molecule level, without the need of chemical modifications and in conditions that mimic an in-vivo state.

The project will deliver a unique bioanalytical tool that will make key contributions to the fundamental understanding of biomolecular interactions, which is needed in basic research as well as in the pharmaceutical industry.

Meccanismo di finanziamento

HORIZON-AG-UN - HORIZON Unit Grant

Coordinatore

FYZIKALNI USTAV AV CR V.V.I
Contribution nette de l'UE
€ 166 278,72
Indirizzo
Na slovance 1999/2
182 21 Praha 8
Cechia

Mostra sulla mappa

Regione
Česko Praha Hlavní město Praha
Tipo di attività
Higher or Secondary Education Establishments
Collegamenti
Contributo UE
Nessun dato