TY - JOUR
T1 - Mouse retinal cell behaviour in space and time using light sheet fluorescence microscopy
AU - Prahst, Claudia
AU - Ashrafzadeh, Parham
AU - Mead, Thomas
AU - Figueiredo, Ana
AU - Chang, Karen
AU - Richardson, Douglas
AU - Venkatraman, Lakshmi
AU - Richards, Mark
AU - Russo, Ana Martins
AU - Harrington, Kyle
AU - Ouarne, Marie
AU - Pena, Andreia
AU - Chen, Dong Feng
AU - Claesson-Welsh, Lena
AU - Cho, Kin Sang
AU - Franco, Cláudio
AU - Bentley, Katie
N1 - Funding Information:
for aiding with 3D drawing of tuft knots. C.P and K.B. were supported by funding from
Funding Information:
K.B. and L.C.W. were supported by a grant
Funding Information:
supported by The Francis Crick Institute, which receives its core funding from Cancer
Funding Information:
P.A. was additionally supported by a travel grant from The Royal Swedish Academy of
Funding Information:
L.V. was funded by the Victor A. McKusick Fellowship from the Marfan Foundation and
Funding Information:
01), and BIDMC. K. B and P.A were supported by The Kjell and Märta Beijer Foundation.
Funding Information:
were supported by funding from Harvard Catalyst| The Harvard Clinical and Translational Science Center (National Center for Research Resources and the National Center for Advancing Translational Sciences, National Institutes of Health Award UL1 TR001102), the NEI (1R21EY027067-01), and BIDMC. K. B and P.A were supported by The Kjell and M?rta Beijer Foundation. P.A. was additionally supported by a travel grant from The Royal Swedish Academy of Sciences (Kungl. Vetenskaps-Akademien). K.B. and L.C.W. were supported by a grant from the Knut and Alice Wallenberg foundation (KAW 2015.0030). K.B. and T.M. were supported by The Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001751), the UK Medical Research Council (FC001751), and the Wellcome Trust (FC001751). L.V. was supported by a Victor A. McKusick fellowship from the Marfan Society. M.R. was supported by an EMBO fellowship (ALTF 2016-923). K.I.H. was supported by institutional training grant T32 HL07893 from the NHLBI of the NIH. L.V. was funded by the Victor A. McKusick Fellowship from the Marfan Foundation and BIDMC. D.F.C supported by EY025259, Lions Foundation, and NEI core grant P30 EY03790. K-S Cho: EY027067. C.A.F was supported by European Research Council starting grant (679368), the Funda??o para a Ci?ncia e a Tecnologia funding (grants: IF/00412/2012; PRECISE-LISBOA-01-0145-FEDER-016394; and a grant from the Fondation Leducq (17CVD03).
Funding Information:
Wellcome Trust (FC001751). L.V. was supported by a Victor A. McKusick fellowship from
Funding Information:
the Marfan Society. M.R. was supported by an EMBO fellowship (ALTF 2016-923). K.I.H.
Funding Information:
was supported by institutional training grant T32 HL07893 from the NHLBI of the NIH.
Publisher Copyright:
© 2020, eLife Sciences Publications Ltd. All rights reserved.
PY - 2020/2
Y1 - 2020/2
N2 - 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 ‟knottedˮ morphology to patholo gical 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.
AB - 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 ‟knottedˮ morphology to patholo gical 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.
UR - http://www.scopus.com/inward/record.url?scp=85083623069&partnerID=8YFLogxK
U2 - 10.7554/eLife.49779
DO - 10.7554/eLife.49779
M3 - Article
C2 - 32073398
AN - SCOPUS:85083623069
SN - 2050-084X
VL - 9
JO - eLife
JF - eLife
M1 - e49779
ER -