Mouse retinal cell behaviour in space and time using light sheet fluorescence microscopy

eLife

Kyle Harrington¹  Claudia Prahst¹  Lakshmi Venkaraman²  Katie Bentley³  Douglas Richardson⁴  Claudio A Franco⁵  Ana Figueiredo⁵  Marie Ouarné⁵  Ana Martins Russo⁵  Andreia Pena⁵  Dong Feng Chen⁶  Karen Chang⁶  Kin-Sang Cho⁷  Parham Ashrafzadeh⁸  Mark Richards⁸  Lena Claesson-Welsh⁸  Thomas Mead⁹ 

[1]Center for Vascular Biology Research and Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, United States

[2]Center for Vascular Biology Research and Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, United States

[3]The Beijer Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden

[4]Center for Vascular Biology Research and Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, United States

[5]The Beijer Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden

[6]The Francis Crick Institute, London, United Kingdom

[7]Department of Informatics, Faculty of Natural and Mathematical Sciences, Kings College London, London, United Kingdom

[8]Biomedical Engineering Department, Boston University, Boston, United States

[9]Harvard Center for Biological Imaging, Department of Molecular and Cellular Biology, Harvard University, Cambridge, United States

[10]Instituto de Medicina Molecular, Lisbon, Portugal

[11]Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, United States

[12]Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, United States

[13]Geriatric Research Education and Clinical Center, Office of Research and Development, Edith Nourse Rogers Memorial Veterans Hospital, Bedford, United States

[14]The Beijer Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden

[15]The Francis Crick Institute, London, United Kingdom

[16]Department of Informatics, Faculty of Natural and Mathematical Sciences, Kings College London, London, United Kingdom

关键词: confocal microscopy;    lightsheet microscopy;    mouse retina;    angiogenesis;    retinopathy of prematurity;    neurovascular;    Mouse;   

DOI  :  10.7554/eLife.49779

来源: publisher

【 摘 要 】

As the general population ages, more people are affected by eye diseases, such as retinopathies. It is therefore critical to improve imaging of eye disease mouse models. Here, we demonstrate that 1) rapid, quantitative 3D and 4D (time lapse) imaging of cellular and subcellular processes in the mouse eye is feasible, with and without tissue clearing, using light-sheet fluorescent microscopy (LSFM); 2) flat-mounting retinas for confocal microscopy significantly distorts tissue morphology, confirmed by quantitative correlative LSFM-Confocal imaging of vessels; 3) LSFM readily reveals new features of even well-studied eye disease mouse models, such as the oxygen-induced retinopathy (OIR) model, including a previously unappreciated ‘knotted’ morphology to pathological vascular tufts, abnormal cell motility and altered filopodia dynamics when live-imaged. We conclude that quantitative 3D/4D LSFM imaging and analysis has the potential to advance our understanding of the eye, in particular pathological, neurovascular, degenerative processes.

【 授权许可】

CC BY