Periodic Reporting for period 2 - PHOREVER (PHOtonic integrated OCT-enhanced flow cytometry for canceR and cardiovascular diagnostics enabled by Extracellular VEsicles discRimination)
Período documentado: 2024-07-01 hasta 2025-06-30
PHOREVER is a Research and Innovation Action (RIA) funded by the European Commission under the Horizon Europe programme, launched on January 1st, 2023 [M01]. The project aims to develop an innovative, compact, and miniaturized point-of-care (PoC) diagnostic device based on photonic integrated circuits (PICs) and microfluidic chips, to enable the reliable detection of extracellular vesicles (EVs) down to 80 nm, to identify EVs carrying disease-specific proteins on their surface, and to measure their concentration in human blood samples.
The PHOREVER multi-sensing platform integrates three complementary sensing modalities:
• Flow cytometry (FCM): for particle detection and classification.
• Fluorescence sensing: for biomarker identification using fluorescent antibodies.
• Optical coherence tomography (OCT): for micro-imaging and noise reduction.
The system architecture relies on two PICs and three microfluidic chips, ensuring both high functionality and compactness.
Within the framework of the project, two specific use cases will be investigated. The first focuses on pancreatic cancer, aiming to monitor disease progression, assess the risk of metastasis, and evaluate treatment efficacy. The second use case addresses stroke, with emphasis on rapid diagnosis and classification of stroke types. In both cases, the platform’s output will be processed using advanced data analysis tools powered by AI algorithms, enabling the interpretation of complex datasets and their correlation with medical records.
The PHOREVER multi-sensing platform integrates three complementary sensing modalities:
• Flow cytometry (FCM): for particle detection and classification.
• Fluorescence sensing: for biomarker identification using fluorescent antibodies.
• Optical coherence tomography (OCT): for micro-imaging and noise reduction.
The system architecture relies on two PICs and three microfluidic chips, ensuring both high functionality and compactness.
Within the framework of the project, two specific use cases will be investigated. The first focuses on pancreatic cancer, aiming to monitor disease progression, assess the risk of metastasis, and evaluate treatment efficacy. The second use case addresses stroke, with emphasis on rapid diagnosis and classification of stroke types. In both cases, the platform’s output will be processed using advanced data analysis tools powered by AI algorithms, enabling the interpretation of complex datasets and their correlation with medical records.
During the second reporting period, the consortium advanced significantly across all technical and scientific work packages, consolidating system design, delivering first-generation components, and preparing the ground for full system integration and validation.
WP1: Project management activities proceeded smoothly, ensuring effective coordination, monitoring of risks, and timely submission of deliverables. Collaboration across partners remained strong, with regular meetings and technical exchanges supporting coherent progress throughout all WPs.
WP2: The overall architecture of the PHOREVER multi-sensing platform was finalised, with designs of the photonic, fluidic, and electronic subsystems consolidated and reported. Plasma samples from biobanks were analysed, identifying EV size ranges of diagnostic relevance and integrating them into AI-driven pipelines. A benchtop OCT system and validated scattering libraries were established as reference tools. Algorithms for OCT reconstruction and FCM operation advanced, while AI-based diagnostic frameworks achieved strong accuracy in stroke detection and showed promising results for pancreatic cancer biomarker identification.
WP3: The first generation of FCM and OCT photonic integrated circuits (PICs) was fabricated on the TriPleX platform, incorporating novel through-hole structures for fluidic integration. A second-generation OCT design was delivered, while optimised microlens designs and 3D printing with sub-micron accuracy were successfully demonstrated. Subcomponent characterisation confirmed strong optical performance, with remaining deviations addressed through design updates for future iterations. First-generation control electronics for both modalities were designed, fabricated, and partially validated, including multi-channel detection boards and modular acquisition units.
WP4: Key modules of the fluidic cartridge were advanced, including multilayer blood filtration to deliver platelet-free plasma, tangential flow filtration for antibody removal, and hydrodynamic focusing achieving narrow, stable streams. A fully stacked prototype cartridge was manufactured, though integration challenges led to the definition of a simplified, de-risked configuration. This new pathway improves alignment, robustness, and serviceability, and will guide the design of the final disposable unit.
WP5 : The consortium implemented advanced processes for packaging and assembly, achieving vertical stacking of FCM and OCT PICs with high precision and integrating fibre arrays capable of handling high optical power. A modular packaging concept was defined, providing flexibility for assembly, maintenance, and upgrades.
WP6: Two disease-specific biobanks were established, one for pancreatic cancer and one for stroke, together comprising several hundred samples. Standardised pre-analytical workflows for EV analysis were implemented and validated, ensuring reproducibility and reliability of results. In vitro EVs from relevant cell lines were also collected, supporting clinical correlation and biomarker discovery.
WP7: A preliminary market outlook and business case were completed, focusing on pancreatic cancer, stroke, and potential additional applications. The analysis outlined possible adoption scenarios, indicative market sizes, and pathways to cost-effective production. In parallel, an updated IPR review confirmed no new risks to the freedom to operate. D/C activities were also expanded, with over 1900 website visitors and 787 social media followers.
WP8/9: Ethics requirements were continuously monitored, with all clinical and data-handling activities performed under approved protocols and GDPR-compliant frameworks.
WP1: Project management activities proceeded smoothly, ensuring effective coordination, monitoring of risks, and timely submission of deliverables. Collaboration across partners remained strong, with regular meetings and technical exchanges supporting coherent progress throughout all WPs.
WP2: The overall architecture of the PHOREVER multi-sensing platform was finalised, with designs of the photonic, fluidic, and electronic subsystems consolidated and reported. Plasma samples from biobanks were analysed, identifying EV size ranges of diagnostic relevance and integrating them into AI-driven pipelines. A benchtop OCT system and validated scattering libraries were established as reference tools. Algorithms for OCT reconstruction and FCM operation advanced, while AI-based diagnostic frameworks achieved strong accuracy in stroke detection and showed promising results for pancreatic cancer biomarker identification.
WP3: The first generation of FCM and OCT photonic integrated circuits (PICs) was fabricated on the TriPleX platform, incorporating novel through-hole structures for fluidic integration. A second-generation OCT design was delivered, while optimised microlens designs and 3D printing with sub-micron accuracy were successfully demonstrated. Subcomponent characterisation confirmed strong optical performance, with remaining deviations addressed through design updates for future iterations. First-generation control electronics for both modalities were designed, fabricated, and partially validated, including multi-channel detection boards and modular acquisition units.
WP4: Key modules of the fluidic cartridge were advanced, including multilayer blood filtration to deliver platelet-free plasma, tangential flow filtration for antibody removal, and hydrodynamic focusing achieving narrow, stable streams. A fully stacked prototype cartridge was manufactured, though integration challenges led to the definition of a simplified, de-risked configuration. This new pathway improves alignment, robustness, and serviceability, and will guide the design of the final disposable unit.
WP5 : The consortium implemented advanced processes for packaging and assembly, achieving vertical stacking of FCM and OCT PICs with high precision and integrating fibre arrays capable of handling high optical power. A modular packaging concept was defined, providing flexibility for assembly, maintenance, and upgrades.
WP6: Two disease-specific biobanks were established, one for pancreatic cancer and one for stroke, together comprising several hundred samples. Standardised pre-analytical workflows for EV analysis were implemented and validated, ensuring reproducibility and reliability of results. In vitro EVs from relevant cell lines were also collected, supporting clinical correlation and biomarker discovery.
WP7: A preliminary market outlook and business case were completed, focusing on pancreatic cancer, stroke, and potential additional applications. The analysis outlined possible adoption scenarios, indicative market sizes, and pathways to cost-effective production. In parallel, an updated IPR review confirmed no new risks to the freedom to operate. D/C activities were also expanded, with over 1900 website visitors and 787 social media followers.
WP8/9: Ethics requirements were continuously monitored, with all clinical and data-handling activities performed under approved protocols and GDPR-compliant frameworks.
The PHOREVER platform is expected to deliver a next-generation multi-sensing solution that integrates flow cytometry, fluorescence, and optical coherence tomography into a single compact and energy-efficient device. This unique combination will enable more accurate detection and characterisation of EVs, addressing unmet clinical needs in the early diagnosis, monitoring, and treatment assessment of diseases such as pancreatic cancer and stroke.
From the work progress achieved within Period 2, a particular strength of the project has been shown to lie in the fluidic unit. The potential of the pre-analytical stage together with the final fluidic cartridge is very high. The system can process plasma obtained directly from whole blood via the integrated filtration module. All key preparation steps — plasma filtering, dilution to the appropriate particle concentration, staining with antibodies, and removal of unbound antibodies — can be performed automatically within the cartridge. This automation minimises personnel effort, reduces handling errors, and improves reproducibility, placing PHOREVER clearly beyond the current state of the art.
At the industrial level, the project advances Europe’s leadership in silicon nitride PICs, hybrid integration, and micro-optofluidic packaging, strengthening the EU photonics manufacturing value chain. To ensure further uptake and eventual commercialisation, PHOREVER will require the completion of the performance evaluation of all individual units, confirmation that they meet the defined requirements, and the delivery of fully integrated demonstrators supported by rigorous system-level testing and validation. This step is essential to guarantee reliability, clinical readiness, and industrial scalability, thereby unlocking the platform’s transformative potential in healthcare and beyond.
From the work progress achieved within Period 2, a particular strength of the project has been shown to lie in the fluidic unit. The potential of the pre-analytical stage together with the final fluidic cartridge is very high. The system can process plasma obtained directly from whole blood via the integrated filtration module. All key preparation steps — plasma filtering, dilution to the appropriate particle concentration, staining with antibodies, and removal of unbound antibodies — can be performed automatically within the cartridge. This automation minimises personnel effort, reduces handling errors, and improves reproducibility, placing PHOREVER clearly beyond the current state of the art.
At the industrial level, the project advances Europe’s leadership in silicon nitride PICs, hybrid integration, and micro-optofluidic packaging, strengthening the EU photonics manufacturing value chain. To ensure further uptake and eventual commercialisation, PHOREVER will require the completion of the performance evaluation of all individual units, confirmation that they meet the defined requirements, and the delivery of fully integrated demonstrators supported by rigorous system-level testing and validation. This step is essential to guarantee reliability, clinical readiness, and industrial scalability, thereby unlocking the platform’s transformative potential in healthcare and beyond.