Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Imaging the structure and dynamic activity of retinal microglia and macrophage-like cells in the living human eye

Yuhua Rui, Daniel M.W. Lee, Min Zhang, Valerie C Snyder, Rashmi Raghuraman, Elena Gofas-Salas, Pedro Mecê, Sanya Yadav, Pavan Tiruveedhula, Kate Grieve, José-Alain Sahel, Marie-Hélène Errera, Ethan A. Rossi
doi: https://doi.org/10.1101/2022.04.30.490173
Yuhua Rui
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
2Eye Center of Xiangya Hospital, Central South University; Hunan Key Laboratory of Ophthalmology; Changsha, Hunan, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Daniel M.W. Lee
3Department of Bioengineering, University of Pittsburgh Swanson School of Engineering, Pittsburgh, PA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Min Zhang
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Valerie C Snyder
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Rashmi Raghuraman
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Elena Gofas-Salas
4INSERM, CNRS, Institut de la Vision, Sorbonne Universite, Paris, Île-de-France, France
5INSERM-DGOS CIC 1423, Centre Hospitalier National d’Ophtalmologie des Quinze-Vingts, Paris, Île-de-France, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Pedro Mecê
6DOTA, ONERA, Universite Paris-Saclay, Saint-Aubin, Île-de-France, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Sanya Yadav
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Pavan Tiruveedhula
6DOTA, ONERA, Universite Paris-Saclay, Saint-Aubin, Île-de-France, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kate Grieve
4INSERM, CNRS, Institut de la Vision, Sorbonne Universite, Paris, Île-de-France, France
5INSERM-DGOS CIC 1423, Centre Hospitalier National d’Ophtalmologie des Quinze-Vingts, Paris, Île-de-France, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
José-Alain Sahel
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Marie-Hélène Errera
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ethan A. Rossi
1Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
4INSERM, CNRS, Institut de la Vision, Sorbonne Universite, Paris, Île-de-France, France
7School of Optometry, University of California, Berkeley, CA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: rossiea@pitt.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

Purpose We recently showed how a refined sequential detection pattern and image processing pipeline for multi-offset adaptive optics scanning light ophthalmoscopy (AOSLO) can increase the contrast of weakly scattering inner retinal structures, including microglia. However, sequential detection was still time-consuming, preventing dynamics from being monitored over short intervals (< 3 mins). Here we show that simultaneous fiber-bundle (FB) detection can overcomes this limitation to reveal the structure and dynamic activity of microglia and macrophage-like cells in healthy and diseased retinae.

Methods We designed and implemented a custom 7-fiber optical FB with one central confocal fiber and six larger fibers for multi-offset detection in AOSLO at a single focal plane. We imaged the ganglion cell layer at several locations at multiple timepoints (from minutes to weeks) in 8 healthy participants and in 4 patients with ocular infections or inflammation, including ocular syphilis and posterior uveitis. Microglia and immune cells were manually segmented to quantify cell morphometry and motility.

Results Fiber-bundle detection reduced single acquisition time to 20-30 seconds, enabling imaging over larger areas and monitoring of dynamics over shorter intervals. Presumed microglia in healthy retinas had an average diameter of 12.8 μm and with a spectrum of morphologies including circular cells and elongated cells with visible processes. We also detected the somas of putative macroglia, potentially astrocytes, near the optic nerve head. Microglia moved slowly in normal eyes (0.02μm/sec, on average) but speed increased in patients with active infections or inflammation (up to 2.37μm/sec). Microglia activity was absent in a patient with chronic uveitis that was quiescent but apparent over short intervals in an active uveitis retina. In a patient with ocular syphilis imaged at multiple timepoints during treatment, macrophage-like cells containing granular internal structures were seen. Decreases in the quantity and motility of these immune cells were correlated with improvements to vision and other structural and systemic biomarkers.

Conclusions FB-AOSLO enable simplified optical setup with easy alignment and implementation. We imaged the fine-scale structure and dynamics of microglia and macrophage-like cells during active infection and inflammation in the living eye for the first time. In healthy eyes we also detected putative glia cell near the optic nerve head. FB-AOSLO offers promise as a powerful tool for detecting and monitoring retinal inflammation and infection in the living eye over short response to treatment.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted May 01, 2022.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Imaging the structure and dynamic activity of retinal microglia and macrophage-like cells in the living human eye
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Imaging the structure and dynamic activity of retinal microglia and macrophage-like cells in the living human eye
Yuhua Rui, Daniel M.W. Lee, Min Zhang, Valerie C Snyder, Rashmi Raghuraman, Elena Gofas-Salas, Pedro Mecê, Sanya Yadav, Pavan Tiruveedhula, Kate Grieve, José-Alain Sahel, Marie-Hélène Errera, Ethan A. Rossi
bioRxiv 2022.04.30.490173; doi: https://doi.org/10.1101/2022.04.30.490173
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Imaging the structure and dynamic activity of retinal microglia and macrophage-like cells in the living human eye
Yuhua Rui, Daniel M.W. Lee, Min Zhang, Valerie C Snyder, Rashmi Raghuraman, Elena Gofas-Salas, Pedro Mecê, Sanya Yadav, Pavan Tiruveedhula, Kate Grieve, José-Alain Sahel, Marie-Hélène Errera, Ethan A. Rossi
bioRxiv 2022.04.30.490173; doi: https://doi.org/10.1101/2022.04.30.490173

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Bioengineering
Subject Areas
All Articles
  • Animal Behavior and Cognition (4383)
  • Biochemistry (9599)
  • Bioengineering (7092)
  • Bioinformatics (24864)
  • Biophysics (12615)
  • Cancer Biology (9957)
  • Cell Biology (14354)
  • Clinical Trials (138)
  • Developmental Biology (7949)
  • Ecology (12107)
  • Epidemiology (2067)
  • Evolutionary Biology (15989)
  • Genetics (10925)
  • Genomics (14743)
  • Immunology (9869)
  • Microbiology (23676)
  • Molecular Biology (9485)
  • Neuroscience (50871)
  • Paleontology (369)
  • Pathology (1539)
  • Pharmacology and Toxicology (2683)
  • Physiology (4015)
  • Plant Biology (8657)
  • Scientific Communication and Education (1509)
  • Synthetic Biology (2397)
  • Systems Biology (6436)
  • Zoology (1346)