Removing physiological motion from intravital and clinical functional imaging data

eLife

Yingxiao Wang¹  Max Nobis²  Pauline Mélénec²  Paul Timpson²  Kurt I Anderson²  Yousuf H Mohammed²  Heidi CE Welch²  Imogen Moran²  Tri Giang Phan³  Michael S Roberts³  David Herrmann³  Jennifer P Morton³  James RW Conway³  Astrid Magenau³  Douglas Strathdee³  Sean C Warren³  Thomas R Cox⁴  Claire Vennin⁵ 

[1] Immunology Division, Garvan Institute of Medical Research, 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;Therapeutics Research Centre, Diamantina Institute, Faculty of Medicine, University of Queensland, Woolloongabba, Australia;

关键词: FLIM;    FRET;    motion correction;    intravital microscopy;    multiphoton;   

DOI  :  10.7554/eLife.35800

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown