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Advancing optical nanoscopy by combining event-based sensors with modulation-enhanced single-molecule localization microscopy

Project description

Advancing super-resolution microscopy

Being able to observe how biological processes take place at the molecular level, is central to understanding life. Super-resolution microscopy helps researchers visualise molecular structures with nanometre precision. However, current methods fail to combine imaging speed and spatial accuracy. With the support of the Marie Skłodowska-Curie Actions programme, the ES-FLUX project aims to address this limitation by introducing innovative sensors capable of recording light changes with high precision. This approach captures both rich spatial and temporal information simultaneously, improving resolution without sacrificing throughput. The project will advance both computational analysis and instrumentation, hoping to expand the power of high-resolution biological microscopy.

Objective

Non-invasive, high-resolution imaging of biological systems often requires optical microscopy. Since overcoming the diffraction barrier, optical nanoscopy, or super-resolution microscopy, with up to ~1-10 nm resolution can be used for imaging immobilized or live biological samples. The final frontier of optical nanoscopy is high-throughput, true molecular imaging (~1 nm precision) in dynamic, high-mobility systems. Currently, the most promising nanoscopy methods employ modulation-enhanced localization microscopy. However, these are compromised either in throughput or in spatiotemporal precision. This trade-off is due to a sensor choice focused on rich temporal (realizing high localization precision) or rich spatial (realizing high throughput) information. A novel sensor architecture, event-based sensors, merges these choices by measuring changes in photon flux on a megapixel-sized chip with asynchronously recorded microsecond-precise timestamped events. With ES-FLUX, I will combine patterned sinusoidal illumination patterns with event-based single-molecule localization microscopy. This will break the stalemate between high throughput and high precision in modulation-enhanced localization microscopy. The spatiotemporal single-molecule event patterns will be analysed separately in the temporal and in the spatial domains, and as such ES-FLUX will improve the resolution compared to traditional single-molecule localization microscopy. It paves the way for molecular precision whilst allowing the high experimental throughput associated with full-frame camera acquisition. Further improvements in the computational analysis and optomechanical setup expands ES-FLUX towards single-particle tracking and spectral multiplexing. In conclusion, ES-FLUX merges modulation-enhanced single-molecule localization microscopy with event-based sensor acquisition and as such expands the boundaries of optical nanoscopy.

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Programme(s)

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Topic(s)

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Funding Scheme

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HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Call for proposal

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(opens in new window) HORIZON-MSCA-2024-PF-01

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Coordinator

TECHNISCHE UNIVERSITEIT DELFT
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.

€ 232 916,16
Total cost

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