Highly Integrated Versatile Laser Source enabling two-photon excitation in digital diagnostics and biomedical research

Project Information

HILIGHT

Grant agreement ID: 101135034

DOI

10.3030/101135034

EC signature date 15 November 2023

Start date 1 December 2023

End date 30 November 2027

Funded under

• Digital, Industry and Space

Total cost

No data

EU contribution

€ 3 185 792,64

Coordinated by

III-V LAB
France

Periodic Reporting for period 2 - HILIGHT (Highly Integrated Versatile Laser Source enabling two-photon excitation in digital diagnostics and biomedical research)

Reporting period: 2025-06-01 to 2026-05-31

Summary of the context and overall objectives of the project

As a key enabling technology, photonics will play a central role in transforming sectors such as healthcare and biomedical research. HILIGHT focuses on the development and demonstration of an innovative two-photon excitation fluorescence lifetime imaging microscopy (2ph-FLIM) platform that enables unprecedented image acquisition rates. The project aims to deliver versatile, compact, and cost-effective laser and detection technologies capable of integration into the existing confocal microscopy system developed by Vivascope.
At the core of the project is the development of a laser, developed by III-V Lab and CSEM, capable of generating customizable pulse bursts at high repetition rates (3–6 GHz). This laser source will support 2ph-FLIM imaging at megapixel-per-second rates and is designed for acrydin fluorescent dye. Its performance in terms of pulse energy and peak power is comparable to the state of the art, but with significant improvements in size, energy efficiency, and manufacturing cost. Combined with a custom-designed digital Silicon Photomultiplier (dSiPM) implemented in CMOS technology, this laser will enable high-speed, high-resolution imaging while overcoming existing limitations in data acquisition and processing bandwidth. The detector, developed by FBK, employs a time-gated photon counting strategy that significantly reduces computational complexity, enabling high signal-to-noise ratio (SNR) measurements and efficient lifetime reconstruction.
HILIGHT’s technologies will be validated in two distinct use cases by Vivascope and Brunel University. The first focuses on digital histopathology, where the project seeks to replace traditional staining techniques with near-instantaneous optical biopsy capabilities. By integrating the HILIGHT system into a VivaScope platform, the project will enable rapid intraoperative diagnostics and enhanced pathology workflows. The second use case explores applications in biomedical research, particularly in the areas of single-cell biochemical phenotyping and optogenetics using three-dimensional culture systems. The project will demonstrate how HILIGHT’s ultrafast laser source can be used for confined photoactivation within tissue and how its detection system can support FLIM-based biochemical sensing at photon count rates far exceeding those of current technologies.
Through these innovations, the HILIGHT project aims to overcome the technical and economic barriers that have limited the broader adoption of two-photon microscopy. It will enable a new generation of medical and research imaging tools that are faster, more accurate, and more accessible. By targeting both clinical and research applications, HILIGHT not only addresses current limitations in biomedical imaging but also opens up new market opportunities across academic, clinical, and industrial settings.

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

During the second period the first laser fabrication has been finalised. Lasers have been mounted on carriers and selected based on static measurements at III-V Lab for further dynamic testing and packaging (WP2). Mounted lasers have been further tested at CSEM to validate the models used when making the designs and check if mode-locking could be obtained (WP10).
On the detection side (WP4), the first HILIGHT detector has been fabricated and delivered back to FBK from LFoundry in August 2025. The detector has been successfully tested and characterized, proving to be fully functional and meeting all design specifications. These results have been accepted for publication in a conference article (European Solid-State Electronics Research Conference) and led to the timely submission of Deliverable D4.2 (“SPAD array performance”) on the EU portal in February 2026. Following this, the hardware platform (PCB), firmware, and software have been developed to integrate the detector into the scanning microscopy systems of the partners VivaScope and BUL. The PCB design was submitted for fabrication, and the FPGA firmware and PC software were finalized in April 2026. The deliverable regarding this integrated prototype is currently being finalized. Multiphoton microscopy and imaging prototype developments side (WP5), a detailed design of the VivaScope integration platform has been completed (modular 40 × 40 mm PCB). Laser (SMA) and SPAD (ribbon/USB-C) interfaces specified and validated with CSEM and FBK. Timing and signal-propagation budget were quantified; open items tracked for PCB finalisation. D5.1 "Design specifications and performance requirements" has been delivered. Development of FLIM applications (WP6), RP2 activities focused on microscopy platform preparation, development of simulation and analytical tools, and validation frameworks for future HILIGHT integration. Main outcomes include the public release of the Python-based HILIGHTer Digital Twin and development of FIREFLY, a user-friendly FLIM analysis platform implementing novel Fisher Information-based methods. These tools support instrument optimisation and high-throughput analysis and received positive feedback from both academic and industrial stakeholders. Three journal articles are expected from this work. Experimental validation using HILIGHT hardware in T6.4 was delayed due to laser fabrication issues.
Development of medical and biomedical application use caes (WP7), RP2 activities focused on establishing and testing biological models ahead of hardware availability. In addition to the HPNE cell lines demonstrated in RP1 under WP6, testing was expanded to cardiomyocyte calcium imaging, patient-derived intestinal organoids, and Acridine Orange applications for chromatin analysis. Initial protocols, imaging workflows, and sample preparation methods were successfully established. These systems will support future integration of HILIGHT systems and validate analytical tools developed in WP6. Progress remains preparatory due to delays in hardware delivery. Exploitation and dissemination, open science & data management (WP8), the main action was the successful organisation and delivery of the HILIGHT 2026 Scientific workshop strengthening dissemination, networking, and collaboration with academic and industrial stakeholders. The HILIGHT website and communication channels have been consolidated, with increased volume and regularity of research-driven content, broader coverage of consortium activities, improved cross-linking with partners, and stronger engagement on social platforms such as LinkedIn. Multiple scientific publications based on project results have been finalized or are beeing prepared, with submissions planned throughout 2026, supporting Open Access dissemination and impact generation. Deliverable D8.4 (Task 8.3 – Exploitation, led by VivaScope) has been delivered, covering business cases, TRL assessment, IP and licensing strategy, market analysis, exploitation roadmap, stakeholder engagement plan, preliminary P&L refresh, and risk register.

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)

The HILIGHT project is expected to deliver a breakthrough in biomedical imaging through the development of an ultrafast, compact, and cost-effective two-photon fluorescence lifetime imaging microscopy (2ph-FLIM) system. By combining a novel monolithic pulse burst laser source with a high-speed, low-noise digital detector, HILIGHT will enable real-time, high-resolution imaging suitable for both clinical and research environments. The main expected results include the successful integration of these technologies into the VivaScope confocal microscope platform, demonstrating unparalleled imaging speed (8 Mpx/s), enhanced depth penetration, and high SNR in digital histopathology and biomedical assays. These advances will allow for near-instantaneous tissue diagnostics, drastically reducing the time required for cancer detection and surgical decision-making. The potential impact of HILIGHT spans several domains. Scientifically, it will open new frontiers in digital histology and cell-level biochemical phenotyping, enabling better understanding and monitoring of disease mechanisms. Economically, it promises to reduce healthcare costs by shortening diagnostic cycles, improving treatment precision, and minimizing unnecessary interventions. Socially, it will contribute to improved patient outcomes and well-being by reducing diagnostic delays and psychological stress. Moreover, the project strengthens European technological sovereignty by developing and industrializing advanced semiconductor photonics within the EU, addressing strategic priorities in healthcare innovation, digital transformation, and global competitiveness.