Periodic Reporting for period 1 - flIMAGIN3D (Doctoral Network for a Shared Excellence of Fluorescent Lifetime Imaging Microscopy in Biomedical Applications)
Reporting period: 2023-01-01 to 2024-12-31
Undoubtedly, fluorescence microscopy is a fundamental pillar supporting basic and applied research, particularly in biological and biomedical sciences. To the non-expert, fluorescence microscopy is primarily based on the analysis of fluorescence intensity emitted but specific molecule (fluorophores) within different spectral windows. However, other properties of light can be used for a more quantitative characterization of biological samples. For example, the average time between excitation and emission of light – the fluorescence lifetime – is modulated by changes within the fluorophore microenvironment such as pH, temperature or non-radiative energy transfer. Therefore, by mapping fluorescence lifetime values in space and time, fluorescence lifetime imaging microscopy (FLIM) provides an exquisitely quantitative characterization of the biochemical and biophysical environment of the cell. Therefore, FLIM has been integrated successfully with most imaging modalities, for example, wide-field, confocal, multiphoton microscopes and light-sheet microscopy thus permitting scientists to investigate cellular biochemistry (e.g. signalling, metabolism, molecular machinery) also in living three-dimensional cultures with low invasiveness either using endogenous or exogenous fluorophores. For these reasons, FLIM has been the key enabling technologies underpinning the discovery and characterization of a multitude of fundamental biological mechanisms. Despite its proven disruptive potential in the biomedical and tissue engineering fields, FLIM is still regarded as a specialist tool. Therefore, we propose flIMAGING3D – a doctoral training network based on an internationally competitive consortium of developers, users and industrial collaborators. flIMAGING3D aims to eliminate the technical barriers and the gap in skills that still limit the broader adoption of FLIM.
The fields of Biomedical Science and Tissue Engineering are hugely important in the care of our citizens and the development of new treatments towards relieving rehabilitating diseases and traumas. Now, more than ever, quick and reliable assessment in clinically relevant models is of utmost importance. Therefore, we believe it is urgent that physicists, chemists and biomedical scientists join forces in the development of a robust and advanced user-friendly FLIM-based platform that is widely accessible to all. In flIMAGIN3D, which centres on fluorescent lifetime imaging microscopy applied to biomedical platforms in 3D, ten talented Doctoral Candidates will be recruited, trained and supervised to cohesively establish the most robust, user accessible and powerful FLIM-based platform-never achieved before, and apply it to a wide range of applications in the biological sciences. Strongly coupled and based on secondments in leading suppliers of these technologies, this will strengthen the bridge(s) needed between the various disciplines. This doctoral network will train talented doctoral candidates in the essential theories underpinning FLIM microscopy, and through the individual projects and extensive secondments in this network; elevate their projects and skills, and that of their secondment hosts to research excellence in FLIM flIMAGIN3D addresses a significant knowledge gap in the field of photonics in biology in that biomedical scientists are involved in each and every aspect of development and design, and cognisant of the wide variety of current platforms that require adaptability to such modalities.
Project website:https://www.flimagin3d.com/
The fields of Biomedical Science and Tissue Engineering are hugely important in the care of our citizens and the development of new treatments towards relieving rehabilitating diseases and traumas. Now, more than ever, quick and reliable assessment in clinically relevant models is of utmost importance. Therefore, we believe it is urgent that physicists, chemists and biomedical scientists join forces in the development of a robust and advanced user-friendly FLIM-based platform that is widely accessible to all. In flIMAGIN3D, which centres on fluorescent lifetime imaging microscopy applied to biomedical platforms in 3D, ten talented Doctoral Candidates will be recruited, trained and supervised to cohesively establish the most robust, user accessible and powerful FLIM-based platform-never achieved before, and apply it to a wide range of applications in the biological sciences. Strongly coupled and based on secondments in leading suppliers of these technologies, this will strengthen the bridge(s) needed between the various disciplines. This doctoral network will train talented doctoral candidates in the essential theories underpinning FLIM microscopy, and through the individual projects and extensive secondments in this network; elevate their projects and skills, and that of their secondment hosts to research excellence in FLIM flIMAGIN3D addresses a significant knowledge gap in the field of photonics in biology in that biomedical scientists are involved in each and every aspect of development and design, and cognisant of the wide variety of current platforms that require adaptability to such modalities.
Project website:https://www.flimagin3d.com/
RO.1: To create, characterise, standardise and test cost-effective platforms' adaptability to all of the various user communities, and to enhance existing imaging capabilities.
Activity in this objective has been fervent with significant progress in developing ISO-FLIM. Parallel electrophysiological measurements of cells correlated with FLIM metabolism is ongoing and will be achieved within 2D cultures, with 3D cultures proving to be more challenging. 3D platforms derived from 3D bioinks with oxygen-sensing capabilities are on track.
Activity in this objective has been fervent with significant progress in developing ISO-FLIM (KCL) described in the report of Deliverable 1.2 due in this reporting period.
RO.2: Optimise the data acquisition and user-data interface whereby large amounts of data are quickly and correctly acquired, processed and interpreted under the control of intelligent software.
Initially, a data management plan (DMP) has been generated by all DCs describing the data being generated.
RO.3: Establish a library of tools to act as biosensors, perturbers (‘actuators’) and microenvironments to assess biological activities and processes and interrogate such activity non-invasively and reproducibly.
So far, oxygen probing sensors have been exchanged between the various partners. A full shared list of sensors has been generated across all DCs.
Activity in this objective has been fervent with significant progress in developing ISO-FLIM. Parallel electrophysiological measurements of cells correlated with FLIM metabolism is ongoing and will be achieved within 2D cultures, with 3D cultures proving to be more challenging. 3D platforms derived from 3D bioinks with oxygen-sensing capabilities are on track.
Activity in this objective has been fervent with significant progress in developing ISO-FLIM (KCL) described in the report of Deliverable 1.2 due in this reporting period.
RO.2: Optimise the data acquisition and user-data interface whereby large amounts of data are quickly and correctly acquired, processed and interpreted under the control of intelligent software.
Initially, a data management plan (DMP) has been generated by all DCs describing the data being generated.
RO.3: Establish a library of tools to act as biosensors, perturbers (‘actuators’) and microenvironments to assess biological activities and processes and interrogate such activity non-invasively and reproducibly.
So far, oxygen probing sensors have been exchanged between the various partners. A full shared list of sensors has been generated across all DCs.