June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Consistent Automatic Spectral Signature Recovery of Human retinal pigment epithelium (RPE) Lipofuscin Components and Drusen in Donors with Age-related Macular Degeneration (AMD) using Multi-Excitation Hyperspectral Autofluorescence (AF) Imaging.
Author Affiliations & Notes
  • Neel Dey
    Computer Science and Engineering, New York University, Brooklyn, New York, United States
  • Sungmin Hong
    Computer Science and Engineering, New York University, Brooklyn, New York, United States
  • Yuehong Tong
    Opthalmology, New York University, New York, New York, United States
  • Taariq Mohammed
    Opthalmology, New York University, New York, New York, United States
  • Rainer Heintzmann
    Leibniz Institute of Photonic Technology, Jena, Germany
    Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University, Jena, Germany
  • Martin Hammer
    Opthalmology, University Hospital Jena, Jena, Germany
    Center for Medical Optics and Photonics, University of Jena, Jena, Germany
  • Guido Gerig
    Computer Science and Engineering, New York University, Brooklyn, New York, United States
  • Christine Curcio
    Opthalmology, University of Alabama at Birmingham, Birmingham, Alabama, United States
  • Thomas Ach
    Opthalmology, University Hospital Würzburg, Würzburg, Germany
  • Zsolt Ablonczy
    Ora Inc, Andover, Massachusetts, United States
  • Theodore Smith
    Opthalmology, New York University, New York, New York, United States
  • Footnotes
    Commercial Relationships   Neel Dey, None; Sungmin Hong, None; Yuehong Tong, None; Taariq Mohammed, None; Rainer Heintzmann, None; Martin Hammer, None; Guido Gerig, None; Christine Curcio, None; Thomas Ach, Novartis (R); Zsolt Ablonczy, None; Theodore Smith, None
  • Footnotes
    Support  R01 EY015520 (RTS), R01 EY021470 (RTS), NEI EY06109 (CC), 2014 von Sallmann Prize (CC), EyeSight Foundation of Alabama (CC), Research to Prevent Blindness (CC), Dr. Werner Jackstädt Foundation (TA), Bavarian Research Alliance (TA), IZKF Würzburg (TA).
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 399. doi:
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    • Get Citation

      Neel Dey, Sungmin Hong, Yuehong Tong, Taariq Mohammed, Rainer Heintzmann, Martin Hammer, Guido Gerig, Christine Curcio, Thomas Ach, Zsolt Ablonczy, Theodore Smith; Consistent Automatic Spectral Signature Recovery of Human retinal pigment epithelium (RPE) Lipofuscin Components and Drusen in Donors with Age-related Macular Degeneration (AMD) using Multi-Excitation Hyperspectral Autofluorescence (AF) Imaging.. Invest. Ophthalmol. Vis. Sci. 2017;58(8):399.

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

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Abstract

Purpose : Three main AF spectral signatures of normal human RPE lipofuscin (LF) S1, S2 and melanolipofuscin (MLF) S3, have been described ex vivo (PMID: 27226929), and in patients with AMD, a sensitive and specific non-LF spectral drusen signature SDr (PMID: 277496960). Prior strategies employed user guided algorithms. We characterize the spectral signatures of these yet to be determined families of fluorophore using a robust, unbiased fully automatic spectral decomposition method with 4 excitation wavelengths.

Methods : Seven RPE flat mounts from seven AMD donors underwent 40X field hyperspectral AF imaging at four excitation wavelengths: 436, 480, 505 and 555 nm, with emissions recorded at 10 nm intervals, 420-720 nm. The 4 hyperspectral data sets for each tissue underwent simultaneous blind source separation into spectra and abundance source images using non-negative matrix factorization initialized by the first 4 non-negative singular-triplets from singular value decomposition [Boutsidis et al, Pattern Recognition, 2008].

Results : Four distinct and characteristic AF emission spectra, 3 from RPE and one from drusen, of human AMD donors were recovered from each of seven tissues. For each of the four spectra, the corresponding spectra from each tissue showed consistently reproducible peaks, shapes and abundance images (Fig 1), across all 4 excitations (Fig 2).

Conclusions : Consistent, automated and unbiased spectral signature recovery across 7 RPE tissues support the physical existence of currently unknown human fluorophore families with these signatures that should be identified with imaging mass spectroscopy for better understanding of RPE physiology in health and disease.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

 

Figure 1. Spectra and tissue abundances of human RPE lipofuscin and drusen. Top Row: AF micrographs, 3 RPE flatmounts. Second Row: 4 similar AF spectra (436 nm excitation) from each tissue. Next four rows: Corresponding tissue abundances. Spectrum 2, SDr, localizes to drusen and subRPE deposits. The other spectra localize to the LF compartment.

Figure 1. Spectra and tissue abundances of human RPE lipofuscin and drusen. Top Row: AF micrographs, 3 RPE flatmounts. Second Row: 4 similar AF spectra (436 nm excitation) from each tissue. Next four rows: Corresponding tissue abundances. Spectrum 2, SDr, localizes to drusen and subRPE deposits. The other spectra localize to the LF compartment.

 

Figure 2. Progression of spectra across excitations. RPE spectrum S1 (top) and Drusen spectrum SDr (bottom) from 3 tissues excited at 436, 480, 505 and 555 nm with consistent shapes and peaks.

Figure 2. Progression of spectra across excitations. RPE spectrum S1 (top) and Drusen spectrum SDr (bottom) from 3 tissues excited at 436, 480, 505 and 555 nm with consistent shapes and peaks.

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