Label-Free Imaging of Inflammation at the Level of Single Cells in the Living Human Eye
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Yes
Purpose
Putative microglia were recently detected using adaptive optics ophthalmoscopy in healthy eyes. Here we evaluate the use of nonconfocal adaptive optics scanning light ophthalmoscopy (AOSLO) for quantifying the morphology and motility of presumed microglia and other immune cells in eyes with retinal inflammation from uveitis and healthy eyes.
Methods
Twelve participants were imaged (8 healthy participants and 4 posterior uveitis patients). The Pittsburgh AOSLO imaging system was used with a custom-designed 7-fiber optical fiber bundle for simultaneous confocal and nonconfocal multioffset detection. The inner retina was imaged at several locations at multiple timepoints to generate time-lapse images. Microglia and macrophages were manually segmented from nonconfocal AOSLO images, and their morphological characteristics quantified. Cell soma motion was quantified across time for periods of up to 30 minutes and their speeds were calculated by measuring their displacement over time.
Results
A spectrum of cell morphologies was detected in healthy eyes from circular amoeboid cells to elongated cells with visible processes, resembling activated and ramified microglia, respectively. Average soma diameter was 16.1 ± 0.9 μm. Cell movement was slow in healthy eyes (0.02 μm/sec on average), but macrophage-like cells moved rapidly in some uveitis patients (up to 3 μm/sec). In an eye with infectious uveitis, many macrophage-like cells were detected; during treatment their quantity and motility decreased as vision improved.
Conclusion
In vivo adaptive optics ophthalmoscopy offers promise as a potentially powerful tool for detecting and monitoring inflammation and response to treatment at a cellular level in the living eye.
Conflict of interest
No
1
Last name
ERRERA
Initials of first name(s)
MH
Department
Department of Ophthalmology, University of Pittsburgh School of Medicine Pittsburgh, Pennsylvania
City
Pittsburgh
Country
United States
2
Last name
Rui
Initials of first name(s)
Y
Department
Department of Ophthalmology, University of Pittsburgh School of Medicine Pittsburgh, Pennsylvania
City
Pittsburgh
Country
United States
3
Last name
Min
Initials of first name(s)
Z
Department
Department of Ophthalmology, University of Pittsburgh School of Medicine Pittsburgh, Pennsylvania
City
Pittsburgh
Country
United States
4
Last name
Snyder
Initials of first name(s)
VC
Department
Department of Ophthalmology, University of Pittsburgh School of Medicine Pittsburgh, Pennsylvania
City
Pittsburgh
Country
United States
5
Last name
Rossi
Initials of first name(s)
EA
Department
Department of Ophthalmology, University of Pittsburgh School of Medicine Pittsburgh, Pennsylvania
City
Pittsburgh, PA
Country
United States
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