Intramolecular optical microscopy with sub-nm spatial resolution in single biomolecules and single-protein sequencing

Project Information

IntraMol

Grant agreement ID: 101065980

DOI

10.3030/101065980

Project closed

EC signature date 22 August 2022

Start date 1 January 2023

End date 31 December 2024

Funded under

• Marie Skłodowska-Curie Actions (MSCA)

Total cost

No data

EU contribution

€ 173 847,36

Coordinated by

MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV
Germany

Periodic Reporting for period 1 - IntraMol (Intramolecular optical microscopy with sub-nm spatial resolution in single biomolecules and single-protein sequencing)

Reporting period: 2023-01-01 to 2024-12-31

Summary of the context and overall objectives of the project

Advancements in biomedical research rely heavily on our ability to visualize biological structures at the smallest possible scale. Traditional optical microscopy is limited by the diffraction of light (approx. 200 nm), making it impossible to resolve molecular structures with atomic precision. Super-resolution microscopy can overcome this barrier but it is still limited to around 10-20 nm in resolution. This project overcomes that limitation by developing Ångström-resolution fluorescence microscopy (Resolution Enhancement by Sequential imaging - RESI), a revolutionary technique capable of visualizing biomolecular interactions at a scale previously thought unattainable.

By achieving a localization precision of approximately 1 Ångström (0.1 nanometers), this technique enables researchers to directly observe molecular arrangements, protein interactions, and structural changes in biological systems with unprecedented clarity. Such insights have profound implications for drug discovery, disease diagnostics, and the development of targeted therapies.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The project focused on developing a revolutionary microscopy technique to achieve, for the first time, Ångström resolution with an optical microscope. By leveraging the precise blinking behavior of fluorescent molecules, sequential super-resolution imaging and implementing sophisticated localization algorithms, we achieved a resolution far beyond what was previously possible with standard super-resolution methods.

Key achievements include:
• The direct visualization of biomolecular distances at the atomic scale.
• Demonstration of RESI’s ability to resolve molecular changes in biological systems, offering insights into protein conformations and interactions.
• Application of the technique to biomedical research, showing its potential for studying disease mechanisms at an unprecedented level of detail.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

This work represents a major leap beyond the current state of the art in microscopy. Existing super-resolution techniques, such as STED and DNA-PAINT, achieve resolutions in the nanometer range, but RESI pushes the boundary to the Ångström scale. This advance opens new doors for structural biology and biomedical research, enabling direct visualization of molecular processes that drive health and disease.

To fully leverage RESI’s potential, future developments will focus on broader accessibility and adaptation for live-cell imaging, integration with other imaging modalities, and further refinement for biomedical applications. These advancements will accelerate discoveries in molecular biology, drug development, and personalized medicine.

Ångström-resolution fluorescence microscopy in synthetic nanostructures and biological samples.