Objective
Metabolic dysregulation is a hallmark of numerous diseases. Multiphoton microscopy (MPM) combined with fluorescence lifetime imaging (FLIM) of the metabolic coenzymes NADH and FAD has emerged as a key method for monitoring metabolic processes in live tissue with sub-cellular resolution. However, recent data demonstrates that spectral crosstalk from other cellular components, such as keratin, is substantial. Yet the contributions from endogenous proteins are often neglected, bringing into question the validity of earlier studies. To address this issue, we propose to systematically scrutinize the fluorescence properties of NADH, FAD and keratin in different states and combinations. This will be done by meticulous investigation of the fluorescence lifetime, spectra and related photophysical properties using MPM and time-resolved spectrophotometry under a range of conditions (pH, temperature, viscosity, protein binding), and in combination with computational chemistry. Based on this knowledge, we will develop approaches that can accurately determine the concentration, diffusion and binding state of NADH and FAD even in the presence of spectral crosstalk from keratin. The strategy is to implement phasor-FLIM analysis, fluorescence lifetime correlation spectroscopy (FLCS) and raster lifetime image correlation spectroscopy (RLICS) to separate the fluorescence contributions of bound and unbound NADH and FAD from the protein background and quantify the spatial distribution of each species with unprecedented spatiotemporal resolution. The developed methodologies will be validated by monitoring cellular metabolism in human primary cells and tumour cell lines. This study will solidify our understanding of NADH and FAD autofluorescence in complex cellular environments and substantially expand the analytical toolset by which quantitative information about metabolic dysregulation can be obtained, thereby advancing MPM-FLIM as a research and diagnostic tool for metabolic diseases.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences biochemistry biomolecules proteins
- natural sciences physical sciences optics microscopy
- natural sciences physical sciences optics spectroscopy
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Keywords
Project’s keywords as indicated by the project coordinator. Not to be confused with the EuroSciVoc taxonomy (Fields of science)
Project’s keywords as indicated by the project coordinator. Not to be confused with the EuroSciVoc taxonomy (Fields of science)
Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
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HORIZON.1.2 - Marie Skłodowska-Curie Actions (MSCA)
MAIN PROGRAMME
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Topic(s)
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Funding Scheme
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships
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Call for proposal
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
(opens in new window) HORIZON-MSCA-2022-PF-01
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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.
405 30 Goeteborg
Sweden
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.