Project description DEENESFRITPL A rapid nanofluidic valve enables imaging of in vivo-like single-molecule interactions Imaging the activities of single molecules currently requires chemical modifications or surface immobilisation to aid visualisation. However, biochemical and kinetic aberrations hinder understanding of in vivo functions and processes. With the support of the Marie Skłodowska-Curie Actions programme, the RAVASI project will enhance the capabilities of its groundbreaking nanofluidic-scattering optical microscopy setup with the aim of facilitating the imaging process. Currently, the molecules do not spend enough time in the nanofluidic channel to accommodate a statistically relevant number of interactions. A rapid nanofluidic valve will enable control of confinement and release of relevant molecules to overcome this obstacle. Show the project objective Hide the project objective Objective 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. Fields of science natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural scienceschemical sciencespolymer sciencesnatural sciencesphysical sciencesopticsmicroscopy Programme(s) HORIZON.1.2 - Marie Skłodowska-Curie Actions (MSCA) Main Programme Topic(s) HORIZON-MSCA-2021-PF-01-01 - MSCA Postdoctoral Fellowships 2021 Call for proposal HORIZON-MSCA-2021-PF-01 See other projects for this call Funding Scheme HORIZON-AG-UN - HORIZON Unit Grant Coordinator FYZIKALNI USTAV AV CR V.V.I Net EU contribution € 166 278,72 Address Na slovance 1999/2 182 21 Praha 8 Czechia See on map Region Česko Praha Hlavní město Praha Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Other funding € 0,00
