Fundus Autofluorescence Lifetimes and Spectral Features of Soft Drusen and Hyperpigmentation in Age-Related Macular Degeneration

Hammer, Martin; Schultz, R.A.; Hasan, Somar; Sauer, Lydia; Klemm, Matthias; Kreilkamp, Lukas; Zweifel, Lynn; Augsten, R; Meller, Daniel

doi:10.1167/tvst.9.5.20

Cited by 18 publications

(37 citation statements)

References 51 publications

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“…Although drusen can virtually be nonfluorescent, most of them emit fluorescence at shorter wavelength and with longer decay times than the RPE. Thus far, this study confirms in vivo FLIO results 6 and an earlier investigation performing spectrally resolved FLIM in the histology of one AMD and one control donor eye. 12 Relative to our previous study, 12 in histologic cross-sections sub-REP deposit autofluorescence is not overlaid by that of the RPE, and we had more donor material.…”

Section: Discussionsupporting

confidence: 89%

“…In a recent clinical study, we found one-third of soft drusen to be hyperfluorescent. 6 Investigating refractile drusen, Suzuki et al 31 found a transition from uniform hyper-FAF to a ring of hyper-FAF. Delori et al 29 explained the central hypofluorescence of drusen with a hyperfluorescent annulus by the thinning of the RPE on top of the druse with a translocation to its rim.…”

Section: Discussionmentioning

confidence: 99%

“…FLIO has revealed a wide variety in drusen autofluorescence lifetimes. 3 , 4 , 6 Longer fluorescence lifetimes in some of the drusen 4 , 6 and a general shift toward shorter emission wavelengths compared with the adjacent fundus, 6 however, indicate an independent autofluorescence contribution from the drusen themselves.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, longer lifetimes in the larger sub-RPE deposits seem to contradict our in vivo findings, where we observed no difference in large (soft) drusen compared with their surrounding fundus. 6 However, it has to be kept in mind that this holds for hyperfluorescent drusen in vivo only. Also here we found highly fluorescent sub-RPE deposits ( Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, this clinical FLIO investigation showed spectral differences between the autofluorescence emission of drusen and surrounding retina/RPE. 6 This differences in lifetime and spectra may reflect different chemical composition. Whereas dominating fluorophores in the RPE are retinal-derived compounds, 7 , 8 drusen/sub-RPE deposits contain considerable fractions of lipids, phospholipids, (lipo-)proteins, and minerals.…”

mentioning

confidence: 99%

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Fluorescence Lifetimes and Spectra of RPE and Sub-RPE Deposits in Histology of Control and AMD Eyes

Schultz

Gamage

Messinger

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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Purpose To investigate fluorescence lifetimes as well as spectral characteristics of drusen and RPE autofluorescence in AMD. Methods Fluorescence lifetimes and spectra of five eyes with AMD and nine control eyes were analyzed in cryosections by means of two-photon excited fluorescence at 960 nm. Spectra were detected at 490 to 647 nm. Lifetimes were measured using time-correlated single photon counting in two spectral channels: 500 to 550 nm and 550 to 700 nm. Fluorescence decays over time were approximated by a series of three exponential functions. The amplitude-weighted mean fluorescence lifetime was determined. Results We identified 196 sub-RPE deposits (AMD, n = 76; control, n = 120) and recorded 241 RPE sites. The peak emission wavelength of sub-RPE deposits was significantly green shifted compared with RPE (peak at 570 nm vs. 610 nm), but did not differ between AMD and control donors. Sub-RPE deposits showed considerably longer mean fluorescence lifetimes than RPE (ch1, 581 ± 163 ps vs. 177 ± 25 ps; ch2, 541 ± 125 ps vs. 285 ± 31 ps; P < 0.001). Sub-RPE deposits found in AMD eyes had longer lifetimes than deposits of controls (ch1, 650 ± 167 ps vs. 537 ± 145 ps; ch2, 600 ± 125 ps vs. 504 ± 111 ps; P < 0.001). In AMD eyes, sub-RPE deposits showed a more homogenous autofluorescence distribution and more deposits were larger than 63 µm than in control eyes. Conclusions Ex vivo fluorescence imaging of sub-RPE deposits in cross-sections enables the separation of their autofluorescence from that of over- or underlying structures. Our analysis showed considerable variability of sub-RPE deposit lifetimes but not spectra. This indicates that sub-RPE deposits either consist of a variety of different fluorophores or expose the same fluorophores to different microenvironments.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Fluorescence Lifetimes and Spectra of RPE and Sub-RPE Deposits in Histology of Control and AMD Eyes

Schultz

Gamage

Messinger

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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Characterization of drusen formation in a primary porcine tissue culture model of dry AMD

Shaw,

Tate,

Periasamy

et al. 2024

Molecular Therapy - Methods & Clinical Development

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Fluorescence Lifetimes and Spectra of RPE and sub-RPE Deposits in Histology of Normal and AMD Eyes

Schultz

Gamage

Messinger

et al. 2020

Preprint

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24Purpose: To investigate autofluorescence lifetimes as well as spectral characteristics of drusen and 25 retinal pigment epithelium (RPE) in age-related macular degeneration (AMD). 26Method: Fluorescence lifetimes and spectra of five eyes with AMD and nine control eyes were analyzed 27 in cryosections by means of two-photon excited fluorescence at 960 nm. Spectra were detected at 490 -28 647nm. Lifetime was measured using time-correlated single photon counting in two spectral channels: 29 500-550nm and 550-700nm. The fluorescence decays over time were approximated by a series of three 30 exponential functions and the amplitude-weighted mean fluorescence lifetime τ m was determined. 31Results: 196 sub-RPE deposits were identified (AMD n=76, healthy n=120) and 230 RPE sites recorded. 32The peak emission wavelength of drusen was significantly green-shifted compared to RPE (peak at 33 570nm vs. 610nm), but not different between patients and controls. Drusen showed considerably longer 34 τ m than RPE: (ch1: 581 ± 163 ps vs. 177 ± 25 ps, ch2: 541 ± 125 ps vs. 285 ± 31 ps, p < 0.001). Drusen 35 found in AMD eyes had longer lifetimes than drusen of controls (ch1: 650 ± 167 ps vs. 537 ± 145 ps, ch2: 36 600 ± 125 ps vs. 504 ± 111 ps, p < 0.001). In addition, drusen in AMD eyes showed a more homogenous 37 fluorescence distribution and more drusen were larger than 63µm than in control eyes. 38Conclusions: Ex vivo fluorescence imaging of drusen in cross-sections enables the separation of their 39 autofluorescence from that of over-or underlying structures. Our analysis showed considerable 40 variability of drusen lifetimes but not spectra. This indicates that drusen consist of a variety of different 41 fluorophores or expose the same fluorophores to different microenvironments. Changes in drusen 42 lifetimes could be related to AMD progression.

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Fundus Autofluorescence Lifetimes and Spectral Features of Soft Drusen and Hyperpigmentation in Age-Related Macular Degeneration

Abstract: lifetimes and spectral features of soft drusen and hyperpigmentation in age-related macular degeneration.

Cited by 18 publications

References 51 publications

Fluorescence Lifetimes and Spectra of RPE and Sub-RPE Deposits in Histology of Control and AMD Eyes

Fluorescence Lifetimes and Spectra of RPE and Sub-RPE Deposits in Histology of Control and AMD Eyes

Characterization of drusen formation in a primary porcine tissue culture model of dry AMD

Fluorescence Lifetimes and Spectra of RPE and sub-RPE Deposits in Histology of Normal and AMD Eyes

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