Imperial College London is installing a new microscope that can peer deep into living tissue without cutting it open. This matters because standard microscopes struggle to see inside thick biological samples like organs or tumours. The new system combines multiphoton and confocal imaging with time-resolved fluorescence and label-free detection, allowing researchers to watch molecules interact in real time inside living tissue, organoids, or even inside a living animal. It can also gather metabolic information without adding fluorescent dyes, by using the tissue’s own autofluorescence. If this succeeds, the microscope will become a shared facility at the Hammersmith Hospital campus, open to Imperial researchers and external collaborators. It could accelerate studies in neuroscience, infection, immunity, and cancer biology by revealing how cells behave in their natural three-dimensional environment. The system also includes software-based superresolution and single-molecule spectroscopy, enabling measurements at the subcellular level that are currently impossible with existing equipment at the facility. The project is primarily about building research capacity—it does not promise a direct clinical application, but it will give scientists a tool to ask deeper questions about fundamental biological mechanisms.
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Light microscopy forms a bedrock of modern research in bioscience and medicine, allowing processes to be examined across scales from individual molecules up to living organisms. We propose to establish an imaging platform with a highly advanced confocal and multiphoton microscope to allow the study of events in cellular ensembles, tissues and in intravital preparations with unparalleled precision and penetration depth. The equipment will permit MRC-funded and other researchers to drive research projects understanding the fundamental mechanisms driven by molecules its interactions and dynamics. Ultimately, we aim to translate cutting-edge research findings into benefits for society. This proposal is to provide a new state-of-the-art microscope for Imperial's Facility for Imaging by Light Microscopy (FILM) at the Hammersmith Hospital/White City campus (HH/WCC). After consulting with the Facility Steering committee and asking the users, we realised the need for a high-end multiphoton with confocal (single-photon) capabilities and time-resolved fluorescence lifetime imaging and fluorescence spectroscopy. From the quotes provided by Zeiss, Nikon and Leica, we favour, for technical reasons, capabilities, and FILM facility know-how, the Leica STELLARIS 8 DIVE FALCON multiphoton and confocal with a software-based superresolution microscope. This system allows time-resolved fluorescence imaging and single-molecule spectroscopy dynamics deep inside cellular ensembles, tissue and intravital, and single cellular and subcellular levels. The system will enable us to resolve functional molecular events spatiotemporally in thick specimens, as we will be able to image deep inside the specimens at levels we are currently unable to achieve with other equipment at FILM. Moreover, we will be able to image events without the need to use a fluorescence probe by employing autofluorescence imaging and second harmonic generation signals, ultimately allowing us to gather quantitative metabolic information without using fluorophores. This will all be quantified by employing the phasor analysis and time-resolved fluorescence (FLIM). This microscope will enhance Imperial College research in several MRC strategic priority areas: collaborative research with users, population and systems medicine, molecular and cellular medicine, infections and immunity, neurosciences and mental health, global health research, translational research and methodology development, better methods, better research. Technically, the objectives of this proposal will be to 1) establish a multiuser facility for multiphoton fluorescence and label-free imaging in thick specimens with time-resolved fluorescence imaging and single-molecule fluorescence spectroscopy open to all researchers at Imperial College and external academics or industrial collaborators run and managed by the FILM at Hammersmith Campus. 2) Enhance the research outputs, drive scientific discovery, and advance knowledge in current and future research within MRC remit. 3) Increase the usage of multiphoton imaging by enabling applications of less experienced microscopists by providing expert technical support. 4) Nurture the establishment of collaborations within Imperial College London and with external users in multidisciplinary research, benefiting from multiphoton and time-resolved microscopy. 5) Train new users, including students, research staff, academics and external users, in the application of multiphoton microscopy, label-free imaging, time-resolved fluorescence microscopy and single-molecule fluorescence spectroscopy.
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