Project ID BE-MI2026_26

ThemeBE-MI

Co Supervisor 1A Simon Poland Faculty of Life Sciences & Medicine, School of Cancer & Pharmaceutical Sciences, Comprehensive Cancer CentreEmail

Co Supervisor 1B Klaus Suhling Faculty of Natural, Mathematical & Engineering Sciences, Department of PhysicsEmail

Development of a Novel Microfluidics-Based Fluorescence Lifetime Imaging Flow Cytometer for Live Cell Sorting

“Fluorescence lifetime imaging (FLIM) is a sophisticated microscopy technique that measures the duration of light emission from excited molecules. It provides intricate details about the cellular environment, such as pH, viscosity, and temperature, and identifies various ions and metabolic states within tissues. Combined with Förster resonance energy transfer (FRET), FLIM can also assess protein interactions, essential for understanding complex biological systems. Crucially for high-throughput applications such as preparative cell sorting, FLIM based technologies have not been developed due to the complexity of the technology, large data sizes and analysis procedures.

This project aims to revolutionize high-throughput cell sorting, which is essential in biomedical research and clinical diagnostics. By providing a more detailed analysis of cells and allowing for the sorting of heterogeneous cell populations, this technology can significantly enhance research in areas such as cancer, immunology, and stem cell biology. The student will develop new technology to sort live cells at high speeds based on their fluorescence lifetimes, enhancing the precision of cell sorting and providing detailed spatial and functional information about each cell. Utilising a unique approach the student will capture high-resolution volumetric images of cells flowing through the microfluidic chip and incorporate on-chip real-time analysis to enable high-speed preparative live cell sorting, validating the technology for preparative sorting of live cells.

This will address current limitations in cell sorting technologies by incorporating advanced imaging and real-time data processing, thus paving the way for new insights and applications in life sciences and healthcare.

• Year 1 Develop the foundational FLIM setup and microfluidic components for high-throughput cell sorting.
• Year 2: Enhance real-time image acquisition and integrate rapid data analysis for improved cell sorting accuracy.
• Year 3: Conduct extensive validation tests with diverse cell types to refine and optimize the sorting technology.
• Year 4: Finalize the technology, publish findings, and disseminate the system through workshops and collaborations.”

Representative Publications

• Mai H., Jarman A., Erdogan A. T., Treacy C., Finlayson N., Henderson R. K., & Poland S. P. “Development of a high-speed line-scanning fluorescence lifetime imaging microscope for biological imaging,” Optics Letters 48 (8), 2042-2045 (2023). DOI: 10.1364/OL.482403 pubmed.ncbi.nlm.nih.gov • Mai H., Poland S. P., Della Rocca F. M., Treacy C., Aluko J., Nedbal J., et al. “Flow cytometry visualization and real-time processing with a CMOS SPAD array and high-speed hardware implementation algorithm,” in Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVIII (SPIE 11243, 2020), pp. 31-37. DOI: 10.1117/12.2544759 spiedigitallibrary.org • Poland S. P., Krstajić N., Monypenny J., Coelho S., Tyndall D., Walker R. J., et al. “A high-speed multifocal multiphoton fluorescence lifetime imaging microscope for live-cell FRET imaging,” Biomedical Optics Express 6 (2), 277-296 (2015). DOI: 10.1364/BOE.6.000277 https://opg.optica.org/boe/fulltext.cfm?uri=boe-6-2-277&id=307502——————————————————————————————————————————• Suhling K., French P. M. W., Phillips D. “Time-resolved fluorescence microscopy,” Photochemical & Photobiological Sciences 4 (1), 13 – 22 (2005). DOI: 10.1039/B412924P https://doi.org/10.1039/B412924P • Nedbal J., Mattioli Della Rocca F., Ivanova I. T., Allan A., Graham J., Walker R., Henderson R. K., Suhling K. “A time-correlated single-photon-counting SPAD array camera with a bespoke data-processing algorithm for lightsheet fluorescence lifetime imaging (FLIM) and FLIM videos,” Scientific Reports 14 (1), 7247 (2024). DOI: 10.1038/s41598-024-56122-1 — https://doi.org/10.1038/s41598-024-56122-1 pubmed.ncbi.nlm.nih.gov • Hirvonen L. M., Nedbal J., Almutairi N., Phillips T. A., Becker W., Conneely T., Milnes J., Cox S., Stürzenbaum S., Suhling K. “Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single-photon counting,” Journal of Biophotonics 13 (2), e201960099 (2020). DOI: 10.1002/jbio.201960099 https://doi.org/10.1002/jbio.201960099 pubmed.ncbi.nlm.nih.gov