Groundbreaking hardware is the foundation of the HILIGHT project, but it is our innovative approach to imaging and data analysis that transforms technological potential into real-world impact. This third pillar of our research focuses on developing the methods, protocols, and analytical tools needed to harness the full power of our unique laser and detector, ensuring the results are not only fast and accurate but also clear and insightful for both clinicians and researchers.
The Challenge: A New Paradigm for Imaging
The HILIGHT system operates differently from conventional microscopes. Our laser generates nanosecond-long bursts of high-frequency pulses, and our detector uses a small number of time-gates to capture the fluorescence signal. While this architecture is key to achieving unprecedented speed, applying standard analysis methods would result in a significant loss of performance and information. To unlock the system’s true potential, we must rethink how we excite molecules and how we interpret the light they emit.
Our Innovative Approach: Maximising Information and Clarity
Led by Brunel University London (BUL), our team is creating a suite of advanced methods that work in synergy with the HILIGHT hardware. We are not just collecting data; we are intelligently shaping and interpreting it at every step.
Key Aspects of Our Application Development:
- Optimised Excitation and Detection Schemes: We are using advanced computational techniques and Monte Carlo simulations to determine the most effective imaging strategies. This allows us to precisely configure the laser’s pulse bursts and the detector’s timing gates to maximise the Signal-to-Noise Ratio (SNR) for fluorescence lifetime measurements. By exploiting the versatility we have built into the system, we can ensure we capture the highest quality data possible.
- Advanced Data Analysis and “Digital Staining”: Raw data is transformed into meaningful results through sophisticated analysis. We use established, user-friendly techniques like phasor analysis to provide quantitative data without requiring deep expertise in FLIM. For clinical applications, we use machine learning and colour projection techniques to translate fluorescence and reflectance data into digital images that resemble the traditional H&E (haematoxylin and eosin) stains that pathologists are trained to interpret. This “digital staining” provides enhanced contrast and familiar visualisation, bridging the gap between cutting-edge technology and established clinical workflow.
- Standardisation and Validation: To ensure our results are reliable, replicable, and reproducible, we are developing a comprehensive set of standards. This includes preparing certified fluorescent samples and realistic, 3D bioprinted cell cultures stained with relevant dyes. These standards will be used for multi-centre validation across the consortium, guaranteeing the robustness of the HILIGHT platform.
These innovative methods are the crucial link between our hardware and our ultimate goals. They are what enable the HILIGHT system to deliver on its promise of providing faster, more specific diagnostics and empowering a new wave of biomedical discovery.
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