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

  1. Claudia Prahst  Is a corresponding author
  2. Parham Ashrafzadeh
  3. Thomas Mead
  4. Ana Figueiredo
  5. Karen Chang
  6. Douglas Richardson
  7. Lakshmi Venkaraman
  8. Mark Richards
  9. Ana Martins Russo
  10. Kyle Harrington
  11. Marie Ouarné
  12. Andreia Pena
  13. Dong Feng Chen
  14. Lena Claesson-Welsh
  15. Kin-Sang Cho
  16. Claudio A Franco
  17. Katie Bentley  Is a corresponding author
  1. Beth Israel Deaconess Medical Center and Harvard Medical School, United States
  2. Uppsala University, Sweden
  3. The Francis Crick Institute, United Kingdom
  4. Instituto de Medicina Molecular, Portugal
  5. Harvard Medical School, United States
  6. Harvard University, United States
  7. University of Idaho, United States
  8. Universidade de Lisboa, Portugal
  9. The Francis Crick Institute / Kings College London, United Kingdom
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Cite this article as: eLife 2020;9:e49779 doi: 10.7554/eLife.49779

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.

Article and author information

Author details

  1. Claudia Prahst

    Center for Vascular Biology Research and Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, United States
    For correspondence
    Claudia.prahst@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
  2. Parham Ashrafzadeh

    Department for Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  3. Thomas Mead

    Cellular Adaptive Behaviour Lab, The Francis Crick Institute, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Ana Figueiredo

    Instituto de Medicina Molecular, Lisbon, Portugal
    Competing interests
    The authors declare that no competing interests exist.
  5. Karen Chang

    Department of Opthalmology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Douglas Richardson

    Harvard Center for Biological Imaging, Department of Molecular and Cellular Biology, Harvard University, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Lakshmi Venkaraman

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  8. Mark Richards

    Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  9. Ana Martins Russo

    Instituto de Medicina Molecular, Lisbon, Portugal
    Competing interests
    The authors declare that no competing interests exist.
  10. Kyle Harrington

    College of Art and Architecture, University of Idaho, Idaho, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Marie Ouarné

    Instituto de Medicina Molecular, Lisbon, Portugal
    Competing interests
    The authors declare that no competing interests exist.
  12. Andreia Pena

    Instituto de Medicina Molecular, Lisbon, Portugal
    Competing interests
    The authors declare that no competing interests exist.
  13. Dong Feng Chen

    Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6283-8843
  14. Lena Claesson-Welsh

    Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4275-2000
  15. Kin-Sang Cho

    Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4285-615X
  16. Claudio A Franco

    Instituto de Medicina Molecular - João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2861-3883
  17. Katie Bentley

    Informatics, The Francis Crick Institute / Kings College London, London, United Kingdom
    For correspondence
    katie.bentley@kcl.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9391-659X

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).

Reviewing Editor

  1. Anna Akhmanova, Utrecht University, Netherlands

Publication history

  1. Received: June 29, 2019
  2. Accepted: February 11, 2020
  3. Accepted Manuscript published: February 19, 2020 (version 1)

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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