September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Semi-automated analysis of lipofuscine and melanolipofuscine granules in the RPE using structured illumination microscopy
Author Affiliations & Notes
  • Nil Celik
    Department of Ophthalmology, University Hospital Heidelberg, Heidleberg, Germany
  • Gerrit Best
    Institute of Molecular Biology, Mainz, Germany
  • Florian Schock
    Institute of Molecular Biology, Mainz, Germany
  • Alena Bakulina
    University Medical Center Mannheim, Experimental Radiation Oncology, Mannheim, Germany
  • Gerd U. Auffarth
    Department of Ophthalmology, University Hospital Heidelberg, Heidleberg, Germany
  • Christoph Cremer
    Institute of Molecular Biology, Mainz, Germany
  • Juergen Hesser
    University Medical Center Mannheim, Experimental Radiation Oncology, Mannheim, Germany
  • Stefan Dithmar
    Department of Ophthalmology, Dr. Horst Schmidt Kliniken, Wiesbaden, Germany
  • Footnotes
    Commercial Relationships   Nil Celik, None; Gerrit Best, None; Florian Schock, None; Alena Bakulina, None; Gerd Auffarth, Abbott Medical Optics, Alcon Laboratories, Inc., Alimera Sciences, Inc., Bausch & Lomb Surgical, Carl Zeiss Meditec, Contamac, Glaukos Corporation, Heidelberg Engineering, HumanOptics, Novartis Pharmaceuticals Corporation, Oculentis, Physiol, Power Vision, Rayner Intraocular Lenses Ltd, Technolas (F), Abbott Medical Optics, Alcon Laboratories, Inc., Allergan, Bausch & Lomb Surgical, Bayer Healthcare Pharmaceuticals, Carl Zeiss Meditec, Hoya, HumanOptics, Kowa, Oculentis, OPHTEC, Physiol, Rayner Intraocular Lenses Ltd, Technolas (R), Abbott Medical Optics, Alcon Laboratories, Inc., Allergan, Bausch & Lomb Surgical, Novartis Pharmaceuticals Corporation, Rayner Intraocular Lenses Ltd, Technolas (C); Christoph Cremer, None; Juergen Hesser, None; Stefan Dithmar, None
  • Footnotes
    Support  Supported by Gertrud Kusen Foundation
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 6036. doi:
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      Nil Celik, Gerrit Best, Florian Schock, Alena Bakulina, Gerd U. Auffarth, Christoph Cremer, Juergen Hesser, Stefan Dithmar; Semi-automated analysis of lipofuscine and melanolipofuscine granules in the RPE using structured illumination microscopy. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6036.

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

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Abstract

Purpose : We already introduced a new analyzing tool for lipofuscine granules (LG) in retinal pigment epithelium (RPE) cells in highly resolved 3D-Structured illumination microscopy (SIM) images automatically. In a further analysis we additionally investigated melanolipofuscine granules (MLG).

Methods : 3D-SIM images were examined for LG and MLG using three different excitation wavelengths (488, 568 and 647 nm). Our automated software identifies and quantifies (number, size and position) single granules in RPE flatmounts. The LG and MLG of an 88-year-old donor were examined for the macular region.

Results : From 541 granules, 411 (76.0%) were identified as LG and 130 (24.0%) as MLG. LG and MLG were distributed towards the cell edge.

Conclusions : Highly resolved 3D-images of LG as well as MLG are obtainable with SIM. LG and MLG can be differentiated and characterized regarding their number and position with a semi-automated analysis of the established SIM data. Quantitative characteristics of autofluorescent granules help to understand cellular processes within the RPE.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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