Periodic Reporting for period 1 - ImAge-D (Imaging Ageing Endothelium at the nanoscale - Doctorates)
Okres sprawozdawczy: 2023-10-01 do 2025-09-30
ImAgE-D will train a new generation of Doctoral Candidates (DCs) in the development and application of newly developed high speed and high-resolution imaging tools in biomedical research. The 10 DCs will be cross-pollinated with concepts and skills in physics and biomedicine, in particular in super-resolution imaging, analytical image reconstruction, and optical micro-manipulation methods. These skills will be applied to reveal for the first time the functionality and morphology (below the diffraction limit of light) of living endothelial cells (EC) that present the main barrier between the blood/lymph and all organs and tissues, and how these vital cells change with ageing. These specialised, highly trained and mobile DCs will have greatly enhanced career prospects. The training in novel physical methods with highly relevant experience in the biomedical sciences will allow them to confidently navigate at the interface of academic, clinical and private sector research within Europe and the world at large.
The Key Research Objectives of ImAgE-D are:
1. To develop novel high-speed super-resolution technologies that enable the dynamic visualisation of living ECs and live imaging of pharmaceutical effects on ECs in microfluidic perfusion chambers.
2. To develop open-access tools for SR-SIM and AFM image reconstruction and blue-prints for high-speed, real-time image reconstruction.
3. To extend SR-SIM and AFM based nanoscopy methods to 3D cell culture, in situ tissue studies, and in vivo studies of endothelium.
4. To obtain mechanistic descriptions of small molecule/pharmaceutical/endogenous plasma factor effects using SR-SIM and AFM on living ECs, and the effect of these agents on ageing EC.
5. To obtain a battery of endothelially active compounds that are beneficial for ageing EC, using the tools developed above.
The Key training objectives of ImAgE-D:
1. To produce a cohort of 10 PhD graduates each with a high level of scientific/research expertise and 1-3 publications in the translation of cutting edge nanoscopy methods, plus transferable skills from compulsory courses in English, entrepreneurship, grant/bid writing, communication and project management.
2. To encourage and enable the DCs to be leading players in cutting-edge research projects with researchrelated skills in cutting edge optical methods, i.e. adaptive optics and high-speed optical nanoscopy, and other methods such as AFM.
3. To nurture entrepreneurial skills in the DCs by equipping them with the skills needed to play leading roles in converting knowledge and ideas into innovative new products and services through exposure to SME environments.
4. To produce well-rounded professionals with an expansive portfolio of transferrable skills, which enable them to solve problems and manage projects independently, resulting in a wide perspective of potential career paths and career development in the European marketplace.
5. To create an active DC network, whose members will shape the future of knowledge generation and innovative diagnostics and therapeutics development in Europe
The Key Research Objectives of ImAgE-D are:
1. To develop novel high-speed super-resolution technologies that enable the dynamic visualisation of living ECs and live imaging of pharmaceutical effects on ECs in microfluidic perfusion chambers.
2. To develop open-access tools for SR-SIM and AFM image reconstruction and blue-prints for high-speed, real-time image reconstruction.
3. To extend SR-SIM and AFM based nanoscopy methods to 3D cell culture, in situ tissue studies, and in vivo studies of endothelium.
4. To obtain mechanistic descriptions of small molecule/pharmaceutical/endogenous plasma factor effects using SR-SIM and AFM on living ECs, and the effect of these agents on ageing EC.
5. To obtain a battery of endothelially active compounds that are beneficial for ageing EC, using the tools developed above.
The Key training objectives of ImAgE-D:
1. To produce a cohort of 10 PhD graduates each with a high level of scientific/research expertise and 1-3 publications in the translation of cutting edge nanoscopy methods, plus transferable skills from compulsory courses in English, entrepreneurship, grant/bid writing, communication and project management.
2. To encourage and enable the DCs to be leading players in cutting-edge research projects with researchrelated skills in cutting edge optical methods, i.e. adaptive optics and high-speed optical nanoscopy, and other methods such as AFM.
3. To nurture entrepreneurial skills in the DCs by equipping them with the skills needed to play leading roles in converting knowledge and ideas into innovative new products and services through exposure to SME environments.
4. To produce well-rounded professionals with an expansive portfolio of transferrable skills, which enable them to solve problems and manage projects independently, resulting in a wide perspective of potential career paths and career development in the European marketplace.
5. To create an active DC network, whose members will shape the future of knowledge generation and innovative diagnostics and therapeutics development in Europe
In the period M1 – M24, the focus of the work of the Coordinator (UiT) were the development and signing of the Consortium Agreement, establishment of Supervisory Board and Sub-Committees, supporting and overseeing the recruitment of DCs and financial management of network activities. Per this reporting period all ten doctoral candidates have accepted offers of employment at their respective host laboratories, with a number of them already concluding or having concluded their first secondments.
UiT participant Prof. Lorena Arranz has moved to the University of Oslo (UiO). The DC3 position was subsequently moved and is now hosted by UiO, which has been added as a beneficiary to the consortium. Deliverables coming from DC3 remain the same as previously described in the GA. The GA has been amended accordingly in M1 (October 2023).
In its role as a coordinator, UiT organized:
• Consortium wide kick-off meeting, held on the 28th of June 2023, remotely.
• First consortium wide meeting, held from the 18th to the 21st of March 2024 in Tromsø, Norway.
• Second consortium wide and mid-term meeting, held from the 18th to the 20th of Nov 2024, remotely.
• Third consortium wide meeting, held from the 31st of March to the 04th of April 2025 in Krakow, Poland.
The doctoral candidates were tasked with the presentation of the work carried out within their respective projects, focusing on the results obtained and progress achieved towards the project aim and scope, also providing insights on the next steps and planned actions for the upcoming months. In order to contribute to the further training of the DCs, compulsory scientific and transferable skills courses were offered and held.
The following deliverables were submitted at the time of this reporting period (September 2025):
D1.1 - In situ imaging protocols for endothelium
D1.2 - Fibre-SIM protocols for in situ imaging of endothelium
D2.1 - DCs competent in cell isolation
D2.2 - Simultaneous fibre-SIMAFM protocols
D2.3 - Optimised culture conditions for cells from different organs
D3.1 - Cost-efficient fibre-SIM microscope
D3.2 - Cost-efficient Bessel beam microscope
D3.3 - AFM fibre-SIM integrated platform
D6.1 – Data Management Plan
D6.2 – Sustainability Plan
D6.3 – Supervisory Board of the Network
D6.4 – Subcommittees
D6.5 – Recruitment Plan
D6.6 – Progress Report submitted to the REA covering the first year implementation of the project
D7.1 – Career Development Plans
D7.3 – Training Agenda
D8.1 – ImAgE-D Website up
D8.2 - Plan for the dissemination and exploitation of results, including communication activities
The following milestones were reached at the time of this reporting period (Sept 2025):
Consortium Agreement: CA signed in advance by all participants and submitted to EU – Feb 2024
Planned recruitments completed: Letters of acceptance from all DCs – April 2025
All fellows enrolled in PhD programs: Enrolment letters from respective institutions – April 2025
Program mid-term check: Visit and report by EC officer – Nov 2024
UiT participant Prof. Lorena Arranz has moved to the University of Oslo (UiO). The DC3 position was subsequently moved and is now hosted by UiO, which has been added as a beneficiary to the consortium. Deliverables coming from DC3 remain the same as previously described in the GA. The GA has been amended accordingly in M1 (October 2023).
In its role as a coordinator, UiT organized:
• Consortium wide kick-off meeting, held on the 28th of June 2023, remotely.
• First consortium wide meeting, held from the 18th to the 21st of March 2024 in Tromsø, Norway.
• Second consortium wide and mid-term meeting, held from the 18th to the 20th of Nov 2024, remotely.
• Third consortium wide meeting, held from the 31st of March to the 04th of April 2025 in Krakow, Poland.
The doctoral candidates were tasked with the presentation of the work carried out within their respective projects, focusing on the results obtained and progress achieved towards the project aim and scope, also providing insights on the next steps and planned actions for the upcoming months. In order to contribute to the further training of the DCs, compulsory scientific and transferable skills courses were offered and held.
The following deliverables were submitted at the time of this reporting period (September 2025):
D1.1 - In situ imaging protocols for endothelium
D1.2 - Fibre-SIM protocols for in situ imaging of endothelium
D2.1 - DCs competent in cell isolation
D2.2 - Simultaneous fibre-SIMAFM protocols
D2.3 - Optimised culture conditions for cells from different organs
D3.1 - Cost-efficient fibre-SIM microscope
D3.2 - Cost-efficient Bessel beam microscope
D3.3 - AFM fibre-SIM integrated platform
D6.1 – Data Management Plan
D6.2 – Sustainability Plan
D6.3 – Supervisory Board of the Network
D6.4 – Subcommittees
D6.5 – Recruitment Plan
D6.6 – Progress Report submitted to the REA covering the first year implementation of the project
D7.1 – Career Development Plans
D7.3 – Training Agenda
D8.1 – ImAgE-D Website up
D8.2 - Plan for the dissemination and exploitation of results, including communication activities
The following milestones were reached at the time of this reporting period (Sept 2025):
Consortium Agreement: CA signed in advance by all participants and submitted to EU – Feb 2024
Planned recruitments completed: Letters of acceptance from all DCs – April 2025
All fellows enrolled in PhD programs: Enrolment letters from respective institutions – April 2025
Program mid-term check: Visit and report by EC officer – Nov 2024
• A new concept of a compact interferometric approach to super-resolution structured illumination microscopy (SR-SIM) based on microelectromechanical systems (MEMS) based mirrors was developed and is currently undergoing final realization.
• Completion of refined and reproducible protocols for whole-mount staining of all three vascular beds (arterial, venous, and lymphatic) across multiple organ systems.
• Establishment of a transgenic mouse model with optimized staining conditions for the isolation and subsequent analysis of liver sinusoidal endothelial cells (LSECs) using fiber-based SR-SIM.
• Dynamic visualization of cell oxygenation by FD-FLIM using genetically encoded sensor dUnOFLS.
• Two machine learning-based methods – CNN and Cellpose-SAM – for automated scoring of LSEC fenestrations are now developed and are in their final stages before release to the public, and for publication.
• Atomic force microscopy (AFM) can visualize LSEC fenestrations in liver paraffin sections.
• AFM can visualize fenestrations in liver paraffin sections.
• SR-SIM can can be used to visualize fenestrations in mouse liver tissue.
• Identification of 3 novel (small molecule) compounds with “positive” effects on endothelial ultrastructure, and one with “negative” effects on endothelial ultrastructure.
• Completion of refined and reproducible protocols for whole-mount staining of all three vascular beds (arterial, venous, and lymphatic) across multiple organ systems.
• Establishment of a transgenic mouse model with optimized staining conditions for the isolation and subsequent analysis of liver sinusoidal endothelial cells (LSECs) using fiber-based SR-SIM.
• Dynamic visualization of cell oxygenation by FD-FLIM using genetically encoded sensor dUnOFLS.
• Two machine learning-based methods – CNN and Cellpose-SAM – for automated scoring of LSEC fenestrations are now developed and are in their final stages before release to the public, and for publication.
• Atomic force microscopy (AFM) can visualize LSEC fenestrations in liver paraffin sections.
• AFM can visualize fenestrations in liver paraffin sections.
• SR-SIM can can be used to visualize fenestrations in mouse liver tissue.
• Identification of 3 novel (small molecule) compounds with “positive” effects on endothelial ultrastructure, and one with “negative” effects on endothelial ultrastructure.