Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
Astrid Magenau
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
Yousuf H Mohammed
Therapeutics Research Centre, Diamantina Institute, Faculty of Medicine, University of Queensland, Woolloongabba, Australia
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
Imogen Moran
St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia; Immunology Division, Garvan Institute of Medical Research, Sydney, Australia
Claire Vennin
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
James RW Conway
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
Pauline Mélénec
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia; Immunology Division, Garvan Institute of Medical Research, Sydney, Australia
Michael S Roberts
Therapeutics Research Centre, Diamantina Institute, Faculty of Medicine, University of Queensland, Woolloongabba, Australia; Therapeutics Research Centre, School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia
Paul Timpson
Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
Intravital microscopy can provide unique insights into the function of biological processes in a native context. However, physiological motion caused by peristalsis, respiration and the heartbeat can present a significant challenge, particularly for functional readouts such as fluorescence lifetime imaging (FLIM), which require longer acquisition times to obtain a quantitative readout. Here, we present and benchmark Galene, a versatile multi-platform software tool for image-based correction of sample motion blurring in both time resolved and conventional laser scanning fluorescence microscopy data in two and three dimensions. We show that Galene is able to resolve intravital FLIM-FRET images of intra-abdominal organs in murine models and NADH autofluorescence of human dermal tissue imaging subject to a wide range of physiological motions. Thus, Galene can enable FLIM imaging in situations where a stable imaging platform is not always possible and rescue previously discarded quantitative imaging data.
