Fluorescence lifetime imaging(FLI)is a powerful tool for investigating molecular processes,microenvironmental parameters,and molecular interactions across tissue to(sub-)cellular levels.Despite its established value i...Fluorescence lifetime imaging(FLI)is a powerful tool for investigating molecular processes,microenvironmental parameters,and molecular interactions across tissue to(sub-)cellular levels.Despite its established value in biomedical applications,conventional FLI techniques suffer from long acquisition times,limiting their utility in real-time scenarios like fast biological processes and rapid clinical image-guided interventions.Here,we introduce a novel FLI approach that achieves real-time capability through single-snapshot acquisitions by combining a large-format time-gated SPAD array with dual-gate acquisition capability and a rapid lifetime determination algorithm,thus eliminating time-consuming temporal data collection.We demonstrate this method’s scalability and versatility across challenging biomedical applications,such as fast neural dynamics(microscale),multimodal 3D volumetric FLI of tumor organoids(mesoscale),and FLI-guided surgical procedures using tissue-mimicking phantoms(macroscale).Overall,this new methodology significantly enhances FLI’s temporal and spatial capabilities,enabling rapid dynamic biomedical signal acquisition and seamless integration into clinical workflows,particularly fluorescence-guided surgery.展开更多
基金funding received from the National Institutes of Health under grants R01-CA271371,R01-CA237267,and R01-CA250636from the UK Engineering and Physical Sciences Research Council(grant nos.EP/T00097X/1,EP/T021020/1).DF acknowledges the Royal Academy of Engineering’s support through the Chairs in Emerging Technology Program.
文摘Fluorescence lifetime imaging(FLI)is a powerful tool for investigating molecular processes,microenvironmental parameters,and molecular interactions across tissue to(sub-)cellular levels.Despite its established value in biomedical applications,conventional FLI techniques suffer from long acquisition times,limiting their utility in real-time scenarios like fast biological processes and rapid clinical image-guided interventions.Here,we introduce a novel FLI approach that achieves real-time capability through single-snapshot acquisitions by combining a large-format time-gated SPAD array with dual-gate acquisition capability and a rapid lifetime determination algorithm,thus eliminating time-consuming temporal data collection.We demonstrate this method’s scalability and versatility across challenging biomedical applications,such as fast neural dynamics(microscale),multimodal 3D volumetric FLI of tumor organoids(mesoscale),and FLI-guided surgical procedures using tissue-mimicking phantoms(macroscale).Overall,this new methodology significantly enhances FLI’s temporal and spatial capabilities,enabling rapid dynamic biomedical signal acquisition and seamless integration into clinical workflows,particularly fluorescence-guided surgery.
