August 2019
Volume 60, Issue 11
Open Access
ARVO Imaging in the Eye Conference Abstract  |   August 2019
Visualizing near-infrared autofluorescence from retinal pigment epithelial cells in AMD using multi-wavelength excitation
Author Affiliations & Notes
  • Kari Viljami Vienola
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Min Zhang
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Jose Sahel
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Ethan A Rossi
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
    Department of Bioengineering, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Footnotes
    Commercial Relationships   Kari Vienola, None; Min Zhang, None; Jose Sahel, None; Ethan Rossi, University of Rochester (P)
  • Footnotes
    Support  Supported by a grant from the Edward N. & Della L. Thome Memorial Foundation to José Alain Sahel, NIH CORE Grant P30 EY08098 to the University of Pittsburgh Department of Ophthalmology, the Eye and Ear Foundation of Pittsburgh, and from an unrestricted grant from Research to Prevent Blindness, New York, N.Y., USA.
Investigative Ophthalmology & Visual Science August 2019, Vol.60, PB0183. doi:https://doi.org/
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    • Get Citation

      Kari Viljami Vienola, Min Zhang, Jose Sahel, Ethan A Rossi; Visualizing near-infrared autofluorescence from retinal pigment epithelial cells in AMD using multi-wavelength excitation. Invest. Ophthalmol. Vis. Sci. 2019;60(11):PB0183. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : To visualize determine the efficacy of multi-wavelength excitation for imaging retinal pigmented epithelial cells (RPEs) in vivo using near-infrared autofluorescence (NIRAF) in healthy controls and age-related macular degeneration (AMD) patients.

Methods : NIRAF imaging of the RPE was performed in healthy volunteers and patients diagnosed with AMD using a multi-channel adaptive optics scanning laser ophthalmoscope (AOSLO). Two separate superluminescent diodes (centered at 663 nm and 795 nm) were used for NIRAF excitation and confocal reflectance imaging. The time-averaged optical power at pupil plane was 50 µW for 663 nm and 180 µW for 795 nm with a 1.5 deg. field of view. NIRAF signal was collected in an emission band from 814 to 870 nm. A 2.1 Airy disk diameter pinhole was placed in front of the NIRAF detector and an automated algorithm was used for pinhole positioning to maximize the NIRAF signal. The NIRAF images were co-registered using the corresponding reflectance channel(s) as the reference for eye motion.

Results : Individual RPE cells were successfully imaged in the macula of healthy controls and AMD patients with either 663 nm excitation, 795 nm excitation, or simultaneous excitation with both. Shorter wavelength excitation alone produced a slightly noisier image; however, this may be due to different focusing of the two wavelengths due to chromatic aberration. RPE morphology was similar between the two excitation wavelengths with a darker center and hyper autofluorescent border, consistent with the known hexagonal mosaic of RPE cells.

Conclusions : Each excitation wavelength alone was able to produce NIRAF emission capable of resolving individual RPE cells. Combined excitation resulted in a more robust signal. Ongoing work is aimed at determining whether this emission arises from the same structures within the RPE and how NIRAF excited with these different wavelengths may be altered in AMD. Simultaneous excitation of RPE fluorophores with a combination of wavelengths may improve imaging of individual RPE cells.

This abstract was presented at the 2019 ARVO Imaging in the Eye Conference, held in Vancouver, Canada, April 26-27, 2019.

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