Mouse retinal cell behaviour in space and time using light sheet fluorescence microscopy
- Beth Israel Deaconess Medical Center and Harvard Medical School, United States
- Uppsala University, Sweden
- The Francis Crick Institute, United Kingdom
- Instituto de Medicina Molecular, Portugal
- Harvard Medical School, United States
- Harvard University, United States
- University of Idaho, United States
- Universidade de Lisboa, Portugal
- The Francis Crick Institute / Kings College London, United Kingdom
Abstract
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, neuro-vascular, degenerative processes.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Data has been provided for Figures 3d, e, Figure 4c, Figures 5b,c,d,e, Figures 7d,e,f, Supp. Figures 2c,d and Supp. Figures 5h, i
Article and author information
Author details
Lena Claesson-Welsh
Funding
National Eye Institute (1R21EY027067-01)
- Claudia Prahst
- Katie Bentley
European Research Council (starting grant (679368))
- Claudio A Franco
Fundação para a Ciência e a Tecnologia (grants: IF/00412/2012)
- Claudio A Franco
Fondation Leducq (17CVD03)
- Claudio A Franco
National Eye Institute (EY027067)
- Kin-Sang Cho
Knut och Alice Wallenbergs Stiftelse (KAW 2015.0030)
- Lena Claesson-Welsh
- Katie Bentley
Francis Crick Institute
- Thomas Mead
- Katie Bentley
Fundação para a Ciência e a Tecnologia (PRECISE-LISBOA-01-0145-FEDER-016394)
- Claudio A Franco
Harvard Catalyst (UL1 TR001102)
- Claudia Prahst
- Katie Bentley
Beth Israel Deaconess Medical Center (startup funds)
- Claudia Prahst
- Lakshmi Venkaraman
- Katie Bentley
Kjell och Märta Beijers Stiftelse
- Parham Ashrafzadeh
- Katie Bentley
Marfan Foundation (Victor A McKusick fellowship)
- Lakshmi Venkaraman
European Molecular Biology Organization (ALTF 2016-923 fellowship)
- Mark Richards
National Heart, Lung, and Blood Institute (T32 HL07893)
- Kyle Harrington
National Eye Institute (EY025259)
- Dong Feng Chen
National Eye Institute (P30 EY03790)
- Dong Feng Chen
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. Mice used in experiments at Beth Israel Deaconess Medical Center were held in accordance with Beth Israel Deaconess Medical Center institutional animal care and use committee (IACUC) guidelines. Animal work performed at Uppsala University was approved by the Uppsala University board of animal experimentation. Transgenic mice were maintained at the Instituto de Medicina Molecular (iMM) under standard husbandry conditions and under national regulations.(ethics approval reference C134/14 and C116/15).
Copyright
© 2020, Prahst et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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