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Single-molecule visualization of individual multi-drug efflux pump activity in living bacteria with nanophotonic biochips

Project description

Visualisation of multi-drug efflux pump activity in living bacteria with nanophotonic biochips

Nanoengineered photonic chips hold great promise for bioimaging. Funded by the Marie Skłodowska-Curie Actions programme, the Illuminate-AMR project aims to improve imaging of membrane processes in cells with photonic biochips and study population dynamics of multi-drug efflux pump activity in antimicrobial-resistant bacteria via wide-field 3D nanoscopy. The objectives include development and testing of a new nanophotonic biochip with which to visualise and track membrane bound proteins in real-time under common light microscopes; development of a direct assay for imaging the presence and activity of multi-drug efflux pumps in bacterial membranes at the single-cell level; and comparison of the biochip platform with modern single-molecule imaging approaches.

Objective

Nanophotonic chips hold great promise in imaging by providing molecular-level insights into biological processes occurring in live cells. By tuning their design, it becomes possible to selectively modify light at their surface such that they can be used to study the properties of biomolecules and cells at high resolution near the interface. On-chip decoupling of the excitation and emission light at multiple wavelengths is however a challenge in optical microscopy. The aim of this project is to substantially improve imaging of membrane processes in cells with biochips and then study population dynamics of multi-drug efflux pump activity in antimicrobial resistant bacteria by widefield nanoscopy. The objectives are (i) to develop and test a new nanophotonic biochip with which to visualize and track membrane bound proteins and their activity in single living bacterial cells under common light microscopes; (ii) to develop a direct assay for imaging the presence and activity of multi-drug efflux pumps in bacterial membranes at the single-cell level; and (iii) to implement and compare the biochip platform with modern single-molecule approaches. This imaging platform is based on encoding several precise periodicities to the surface of the biochip in order to diffraction-couple the light to traveling surface waves at multiple resonant wavelengths. This biochip will be a superior alternative to expensive TIRF microscopes that have a limited field of view, and will practically offer an improved evenness of illumination, smaller footprint, and a more accurate knowledge of penetration depth of the evanescent field. The biochip will be used to elucidate how phenotypic heterogeneity in multi-drug efflux pump activity contributes to bacterial survival and thus help improve current antibiotic treatment schemes. Such technology will result in immediate benefits for integrative cellular and molecular biology research communities, and thus has a strong potential for commercialization.

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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2020

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Coordinator

BUNDESANSTALT FUER MATERIALFORSCHUNG UND -PRUEFUNG
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 174 806,40
Address
Unter den Eichen 87
12205 Berlin
Germany

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Region
Berlin Berlin Berlin
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Research Organisations
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 174 806,40
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