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

Feb 19, 2020·
Claudia Prahst
,
Parham Ashrafzadeh
,
Thomas Mead
,
Ana Figueiredo
,
Karen Chang
,
Douglas Richardson
,
Lakshmi Venkaraman
,
Mark Richards
,
Ana Martins Russo
,
Kyle Harrington
,
Marie Ouarné
,
Andreia Pena
,
Dong Feng Chen
,
Lena Claesson-Welsh
,
Kin-Sang Cho
,
Claudio a Franco
,
Katie Bentley
· 0 min read
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, neurovascular, degenerative processes.
Type
Publication
eLife, 9