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

Summary
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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More information & hyperlinks
Web resources: https://cordis.europa.eu/project/id/101032071
Start date: 01-01-2022
End date: 22-05-2024
Total budget - Public funding: 174 806,40 Euro - 174 806,00 Euro
Cordis data

Original description

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.

Status

SIGNED

Call topic

MSCA-IF-2020

Update Date

28-04-2024
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Horizon 2020
H2020-EU.1. EXCELLENT SCIENCE
H2020-EU.1.3. EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions (MSCA)
H2020-EU.1.3.2. Nurturing excellence by means of cross-border and cross-sector mobility
H2020-MSCA-IF-2020
MSCA-IF-2020 Individual Fellowships