Kokona, Despina; Schneider, Nadia; Giannakaki-Zimmermann, Helena; Jovanovic, Joel; Ebneter, Andreas; Zinkernagel, Martin (2017). Noninvasive Quantification of Retinal Microglia Using Widefield Autofluorescence Imaging. Investigative ophthalmology & visual science, 58(4), pp. 2160-2165. Association for Research in Vision and Ophthalmology 10.1167/iovs.16-20916
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Purpose
To validate widefield autofluorescence (AF) in vivo imaging of the retina in mice expressing green fluorescent protein (gfp) in microglia, and to monitor retinal microglia reconstitution in vivo after lethal irradiation and bone marrow transplantation.
Methods
Transgenic Cx3cr1gfp/gfp and wildtype Balb/c mice were used in this study. A confocal scanning laser ophthalmoscope was used for AF imaging with a 55° and a widefield 102° lens. Intrasession reproducibility was assessed for each lens. To investigate reconstitution in vivo, bone marrow from Cx3cr1gfp/gfp mice was used to rescue lethally irradiated wildtype mice. Data were compared to confocal microscopy of retinal flat mounts.
Results
Both the 55° and the 102° lens produced high resolution images of retinal microglia with similar microglia density. However, compared to the 55° lens, the widefield 102° lens captured approximately 3.6 times more microglia cells (1515 ± 123 cells versus 445 ± 76 cells [mean ± SD], for 102° and 55°, respectively, P < 0.001). No statistical difference in the number of gfp positive cells within corresponding areas was observed within the same imaging session. Imaging of microglia reconstitution showed a similar time course compared to flat mount preparations with an excellent correlation between microglia cell numbers in AF and gfp-stained flat mounts (R = 0.92, P < 0.0001).
Conclusions
Widefield AF imaging of mice with gfp expressing microglia can be used to quantify retinal microglia. In vivo microglia counts corresponded very well with ex vivo counts on retinal flat mounts. As such, AF imaging can largely replace ex vivo quantification.
Item Type: |
Journal Article (Original Article) |
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Division/Institute: |
04 Faculty of Medicine > Department of Head Organs and Neurology (DKNS) > Clinic of Ophthalmology |
Graduate School: |
Graduate School for Cellular and Biomedical Sciences (GCB) |
UniBE Contributor: |
Ebneter, Andreas, Zinkernagel, Martin Sebastian |
Subjects: |
600 Technology > 610 Medicine & health |
ISSN: |
0146-0404 |
Publisher: |
Association for Research in Vision and Ophthalmology |
Language: |
English |
Submitter: |
Andreas Ebneter |
Date Deposited: |
14 Aug 2017 10:06 |
Last Modified: |
02 Mar 2023 23:29 |
Publisher DOI: |
10.1167/iovs.16-20916 |
PubMed ID: |
28395300 |
BORIS DOI: |
10.7892/boris.99297 |
URI: |
https://boris.unibe.ch/id/eprint/99297 |
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