Integrated Ultrasound-Enhanced Biophotonic Platform for High-Throughput Studies of Cancer–Immune Cell Mechanobiology

PROJECT OVERVIEW

Mechanobiology, the study of how physical forces and tissue mechanics influence biological function, is emerging as a critical but underexplored layer of regulation in cancer immunology. Within the tumour microenvironment (TME), mechanical cues such as extracellular matrix stiffness, interstitial pressure, and dynamic forces profoundly influence immune cell activation, migration, and effector function. These physical factors can either suppress or enhance immune responses, yet their quantification and measurement are rarely integrated into conventional experimental pipelines. This DPhil project aims to address that gap through the development and application of an integrated ultrasound-enhanced biophotonic imaging platform, specifically designed to investigate immune–cancer mechanobiology from single cells to large-scale populations, with unprecedented spatial and temporal resolution.

At the core of this project is the design and implementation of a second-generation, inverted light-sheet imaging system with real-time ultrasound-based mechanical stimulation. This will allow precise control of the mechanical forces applied to tumour models, enabling investigation of how defined stimuli, such as compression, oscillatory stress, or localised mechanical inputs, shape immune surveillance, T cell activation, and cytotoxic function.

This work builds on a fully developed first-generation light-sheet imaging platform, designed to overcome the field-of-view and speed limitations of commercial imaging systems. This existing system enables high-speed imaging of a 5 × 5 mm area, roughly 100 times larger than typical setups, at millisecond temporal resolution, allowing simultaneous tracking of up to 100,000 individual cells. This unique combination of scale and speed makes it possible to detect and analyse rare, dynamic events in immune cell behaviour within large cell populations. Additionally, this platform features a machine-learning-based image analysis pipeline, optimised for handling terabytes of time-lapse imaging data enabling automated tracking, rare event detection, and high-dimensional analysis of heterogeneous cell populations.

The new platform will be applied to well-characterised antigen-specific tumour–T cell models, including those expressing the 1G4 T-cell receptor (TCR) and OT-1 TCR, which provide powerful systems to dissect T cell engagement, synapse formation, and cytotoxicity under defined mechanical conditions. If successful, the system could also be extended to study patient-derived T cell responses, opening the door to personalised investigations of tumour mechanobiology.

This interdisciplinary project sits at the intersection of photonics, mechanobiology, and cancer immunology. It offers the DPhil student the opportunity to gain hands-on experience in advanced optical engineering, ultrasound stimulation, live-cell imaging, immune functional assays, and computational analysis using machine learning. The project is designed to deliver both technological innovation and fundamental biological insight.

Finally, this work will contribute to a deeper understanding of how mechanical forces regulate immune–tumour interactions at different scales and may inform the design of next-generation immunotherapies and biomaterials, ones that respond not only to molecular signals but also to the physical realities of the tumour microenvironment.

KEYWORDS

Biophysics, Microscopy, Cancer, T Cells, Mechanobiology 

TRAINING OPPORTUNITIES

The prospective DPhil candidate will be involved in a variety of tasks, including the development and operation of the novel ultrasound-integrated light-sheet microscopy platform, and setting up immune functional imaging assays. Additionally, the DPhil candidate will contribute to the analysis of imaging data using an advanced bioimage analysis pipeline developed within the lab. The training will cover microscopy techniques, ultrasound-based stimulation, and immune cell biology, equipping the candidate with both experimental and analytical skills essential to the project.

Supervision will be provided by Dr. Veronika Pfannenstill and Prof. Marco Fritzsche, with additional support from senior postdoctoral researchers and technical staff in the Biophysical Immunology (BPI) Laboratory. The BPI Lab fosters a collaborative and interdisciplinary research environment, with a strong focus on advanced microscopy and immune cell biology. The DPhil candidate will benefit from extensive training opportunities, attend seminars led by world-renowned scientists, and regularly present their work at weekly BPI lab meetings. Further supervision and scientific input will be offered by Dr. Isabela Pedroza-Pacheco and her team, a functional immunology lab with expertise in immune responses to cancer. Their guidance will be particularly valuable in the second phase of the project.

KEY PUBLICATIONS

Fritzsche, M., & Kruse, K. (2024). Mechanical force matters in early T cell activation. Proceedings of the National Academy of Sciences of the United States of America, 121(37), e2404748121. https://doi.org/10.1073/pnas.2404748121

Pfannenstill, V., Barbotin, A., Colin-York, H., & Fritzsche, M. (2021). Quantitative methodologies to dissect immune cell mechanobiology. Cells, 10(4), 851. https://doi.org/10.3390/cells10040851

Dekkers, J. F., Alieva, M., Cleven, A., Keramati, F., Wezenaar, A. K. L., van Vliet, E. J., Puschhof, J., Brazda, P., Johanna, I., Meringa, A. D., Rebel, H. G., Buchholz, M. B., Barrera Román, M., Zeeman, A. L., de Blank, S., Fasci, D., Geurts, M. H., Cornel, A. M., Driehuis, E., Millen, R., … Rios, A. C. (2023). Uncovering the mode of action of engineered T cells in patient cancer organoids. Nature Biotechnology, 41(1), 60–69. https://doi.org/10.1038/s41587-022-01397-w

Moody, M. A., Pedroza-Pacheco, I., Vandergrift, N. A., Choi, C. W., Lloyd, K. E., Parks, R., … Haynes, B. F. (2016). Immune perturbations in HIV-1–infected individuals who make broadly neutralizing antibodies. Science Immunology, 1(1), aag0851. https://doi.org/10.1126/sciimmunol.aag0851

Fritzsche, M. (2020). Thinking multi-scale to advance mechanobiology. Communications Biology, 3, 469. https://doi.org/10.1038/s42003-020-01197-5

THEMES

Cancer, Mechanobiology, Biophysics, Microscopy

CONTACT INFORMATION OF ALL SUPERVISORS

Email: marco.fritzsche@kennedy.ox.ac.uk, isabela.pedroza-pacheco@ndm.ox.ac.uk, veronika.pfannenstill@kennedy.ox.ac.uk