Journal article
Time-resolved autofluorescence imaging of human donor retina tissue from donors with significant extramacular drusen
Investigative ophthalmology & visual science, Vol.53(7), pp.3376-3386
06/08/2012
DOI: 10.1167/iovs.11-8970
PMCID: PMC3390004
PMID: 22511622
Abstract
Time and spectrally resolved measurements of autofluorescence have the potential to monitor metabolism at the cellular level. Fluorophores that emit with the same fluorescence intensity can be discriminated from each other by decay time of fluorescence intensity after pulsed excitation. We performed time-resolved autofluorescence measurements on fundus samples from a donor with significant extramacular drusen. Tissue sections from two human donors were prepared and imaged with a laser scanning microscope. The sample was excited with a titanium-sapphire laser, which was tuned to 860 nm, and frequency doubled by a BBO crystal to 430 nm. The repetition rate was 76 MHz and the pulse width was 170 femtoseconds (fs). The time-resolved autofluorescence was recorded simultaneously in 16 spectral channels (445-605 nm) and bi-exponentially fitted. RPE can be discriminated clearly from Bruch's membrane, drusen, and choroidal connective tissue by fluorescence lifetime. In RPE, bright fluorescence of lipofuscin could be detected with a maximum at 510 nm and extending beyond 600 nm. The lifetime was 385 ps. Different types of drusen were found. Most of them did not contain lipofuscin and exhibited a weak fluorescence, with a maximum at 470 nm. The lifetime was 1785 picoseconds (ps). Also, brightly emitting lesions, presumably representing basal laminar deposits, with fluorescence lifetimes longer than those recorded in RPE could be detected. The demonstrated differentiation of fluorescent structures by their fluorescence decay time is important for interpretation of in vivo measurements by the new fluorescence lifetime imaging (FLIM) ophthalmoscopy on healthy subjects as well as on patients.
Details
- Title: Subtitle
- Time-resolved autofluorescence imaging of human donor retina tissue from donors with significant extramacular drusen
- Creators
- Dietrich Schweitzer - Department of Ophthalmology, University Hospital Jena, Jena, Germany. schweitzer@med.uni-jena.deElizabeth R GaillardJames DillonRobert F MullinsStephen RussellBirgit HoffmannSven PetersMartin HammerChristoph Biskup
- Resource Type
- Journal article
- Publication Details
- Investigative ophthalmology & visual science, Vol.53(7), pp.3376-3386
- Publisher
- United States
- DOI
- 10.1167/iovs.11-8970
- PMID
- 22511622
- PMCID
- PMC3390004
- ISSN
- 0146-0404
- eISSN
- 1552-5783
- Grant note
- EY017451 / NEI NIH HHS R01 EY017451 / NEI NIH HHS
- Language
- English
- Date published
- 06/08/2012
- Academic Unit
- Ophthalmology and Visual Sciences
- Record Identifier
- 9983980048002771
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