Related Experiment Videos
Noninvasive Quantification of Retinal Microglia Using Widefield Autofluorescence Imaging
Despina Kokona1, Nadia Schneider1, Helena Giannakaki-Zimmermann1
1Department of Ophthalmology and Department of Clinical Research, Inselspital, Bern University Hospital, and University of Bern, Switzerland.
Investigative Ophthalmology & Visual Science
|April 11, 2017
Summary
Widefield autofluorescence (AF) imaging effectively quantifies retinal microglia in mice. This in vivo method closely matches ex vivo cell counts, offering a less invasive alternative for microglia research.
Area of Science:
- Ophthalmology
- Immunology
- Cell Biology
Background:
- Retinal microglia play crucial roles in ocular health and disease.
- Accurate quantification of microglia is essential for studying retinal conditions.
- Current methods for microglia quantification can be invasive and time-consuming.
Purpose of the Study:
- To validate widefield autofluorescence (AF) in vivo imaging for assessing retinal microglia in mice.
- To monitor the reconstitution of retinal microglia following bone marrow transplantation.
- To compare the efficacy of widefield AF imaging with traditional ex vivo methods.
Main Methods:
- Utilized transgenic Cx3cr1gfp/gfp mice and wildtype Balb/c mice.
- Employed a confocal scanning laser ophthalmoscope with 55° and widefield 102° lenses for AF imaging.
- Assessed intrasession reproducibility and compared in vivo AF imaging with ex vivo confocal microscopy of retinal flat mounts.
Main Results:
- Both 55° and 102° lenses provided high-resolution images of retinal microglia.
- The widefield 102° lens captured significantly more microglia cells (3.6x) compared to the 55° lens.
- In vivo microglia counts using AF imaging showed excellent correlation (R=0.92) with ex vivo counts from gfp-stained flat mounts.
Conclusions:
- Widefield AF imaging is a validated method for quantifying retinal microglia in vivo.
- This technique accurately reflects ex vivo quantification, potentially replacing it.
- AF imaging offers a powerful tool for monitoring microglia dynamics in research settings.