Fast Infrared Coherent Harmonic Microscopy

Project Information

FAIR CHARM

Grant agreement ID: 101016457

DOI

10.3030/101016457

EC signature date 15 December 2021

Start date 1 January 2022

End date 30 June 2026

Funded under

• INDUSTRIAL LEADERSHIP - Leadership in enabling and industrial technologies - Information and Communication Technologies (ICT)

Total cost

€ 5 944 726,25

EU contribution

€ 5 944 726,25

Coordinated by

UNIVERSITE DE GENEVE
Switzerland

Periodic Reporting for period 2 - FAIR CHARM (Fast Infrared Coherent Harmonic Microscopy)

Reporting period: 2023-07-01 to 2024-12-31

Summary of the context and overall objectives of the project

The EU-funded FAIR CHARM project aims at developing and providing to the research and clinical communities two complementary imaging solutions, SWIM and SLIDE, which transform the capacity to capture in real-time the biological processes as well as the cellular and extracellular structures involved in disease onset and progression. While the SWIM microscope uses novel laser sources in the infrared wavelength region up to 1700 nm to penetrate deeper into biological tissue, the SLIDE microscope pushes the boundaries of fast multiphoton microscopy to imaging rates higher than several thousand images per second. The FAIR CHARM consortium is composed by an interdisciplinary team including photonics innovators for the development of the new instruments, biophotonics experts, and end-users active in the domains of oncology, inflammation, regenerative medicine, and neural signaling willing to address pivotal biomedical questions through the novel technologies developed in the project.

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 Lithuanian laser company Light Conversion made significant modifications to their infrared laser source (CRONUS 3P) to enhance its performance, compactness, and long-term stability. The German partner Miltenyi Biotec focused on extending the wavelength range accessible by their imaging platform, built upon the existing TriM Scope Matrix. Thanks to these modifications, the partners realized the complete integration of the new Light Conversion source within the microscope (dubbed SWIM), both in terms of spectral and optical properties, and software control.
Since summer 2023 research-grade SWIM systems are available to all project partners and installed at the Max Planck Institute in Göttingen and at the University.
Concurrently, the SME Medizinisches Laserzentrum Lübeck GmbH with the support of the University of Lübeck successfully built the first rugged, movable prototype of a microscope that implements SLIDE (diffraction-based) technology for high-speed imaging. The system has already been tested in multiple laboratories of the consortium on relevant samples. Meanwhile, the groups not directly involved in instrument development have made progress in refining and adapting AI-based software analysis tools, in developing ad hoc simulation for heat deposition and dissipation (one the most critical aspects to address when working in this wavelength/power range), in identifying samples and preparation protocols for the biomedical tasks of the project, and in carrying out the characterization of the optical and biophotonic properties of the new systems. Moreover, the partners have contacted and initiated collaborations with clinical experts outside the consortium to further enlarge the scope of the project and for demonstrating and disseminating the diagnostic potential of the new devices. These endeavors have been accompanied by comprehensive dissemination activities involving all project teams, including articles, conference presentations, outreach events, and, notably, the organization of a conference session dedicated to FAIR CHARM at a major European scientific event of 2023.
Since the installation (July 2023) of the first version of the prototype SWIM and SLIDE at the partners location, the consortium has proceeded with their systematic characterization and the assessment of their effect when applied to biological (including in vivo) samples. The former aspect has brought to the continuous optimization of the systems both in terms of performance and in terms of usability by end users. Some of the hardware and software improvements can be considered incremental if considered on their own, but overall, they allow the transfer of the instruments from an optical laboratory environment to realistic bioimaging settings. Some of the upgrades implemented, such as the extended dispersion compensation range of the OPA light source, also align with the broader exploitation of these instruments beyond the FAIR CHARM consortium. By enabling their integration with microscopy platforms from other suppliers, these enhancements further strengthen the impact on the scientific community. Finally, during 2024, the activities of the workpackage focused biomedical challenges have started, including an intense campaign of in vivo measurements within the regenerative medicine axis of the project addressing the detection of nanoparticle-labelled stem cells in rodents in the context of preclinical assays for a cell therapy to counteract Duchenne myopathy.
In response to discussions with the expert EU reviewers, the members of the Advisor Committee, and taking into full account evolving exploitation strategies by key industrial consortium partners, the consortium partners have also expanded the project focus beyond fundamental research. Specifically, they have explored prospective applications of label-free nonlinear imaging to targeted applications using dedicated, smaller footprint, cost-effective platforms. Applications not originally planned in the proposal have been demonstrated to be feasible due to the characteristics of the newly developed systems.

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)

FAIR CHARM groups were able to demonstrate the acquisition of a three-dimensional microscopy movie at 40 volumes/s with the transportable SLIDE system. Furthermore, the collaboration among several project partners led to the detection and imaging of stem cells within untreated whole blood at an imaging speed 200-times higher than that attainable by current cytometry systems. Notably, this scanning speed is too fast to be compatible with the typical lifetimes of fluorophores. Instead, second-harmonic generation has been used for probing the sample. Such an approach holds promise for monitoring cell therapies in preclinical models, assessing their effectiveness, and identifying potential threats such as the presence of cell aggregates in systemic circulation that can prevent their translation. Concerning the SWIM system, the new version of the CRONUS 3P laser by Light Conversion is already at a high TRL level during the first reporting period and it possesses unique characteristics in terms of spectral tunability and long/short-term pulse-stability.
For SWIM, the primary outcome of the thermal assessment is that using a 1 MHz source, instead of the conventional 80 MHz systems, offers a substantial advantage in terms of sensitivity and sample photo-stability. Concerning the exploitation strategy, we would like to emphasize the extension of the dispersion compensation unit of the LCO CRONUS 3P laser following the characterization of the instrument, that will make the system compatible with a broader range of microscopes from multiple suppliers increasing its impact in the field.

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