Chantal Dysli

Quantitative analysis of fluorescence lifetime measurements of the macula using the fluorescence lifetime imaging ophthalmoscope in healthy subjects.

PURPOSE Fundus autofluorescence (FAF) cannot only be characterized by the intensity or the emission spectrum, but also by its lifetime. As the lifetime of a fluorescent molecule is sensitive to its local microenvironment, this technique may provide more information than fundus autofluorescence imaging. We report here the characteristics and repeatability of FAF lifetime measurements of the human macula using a new fluorescence lifetime imaging ophthalmoscope (FLIO). METHODS A total of 31 healthy phakic subjects were included in this study with an age range from 22 to 61 years. For image acquisition, a fluorescence lifetime ophthalmoscope based on a Heidelberg Engineering Spectralis system was used. Fluorescence lifetime maps of the retina were recorded in a short- (498-560 nm) and a long- (560-720 nm) spectral channel. For quantification of fluorescence lifetimes a standard ETDRS grid was used. RESULTS Mean fluorescence lifetimes were shortest in the fovea, with 208 picoseconds for the short-spectral channel and 239 picoseconds for the long-spectral channel, respectively. Fluorescence lifetimes increased from the central area to the outer ring of the ETDRS grid. The test-retest reliability of FLIO was very high for all ETDRS areas (Spearmans ρ = 0.80 for the short- and 0.97 for the long-spectral channel, P < 0.0001). Fluorescence lifetimes increased with age. CONCLUSIONS The FLIO allows reproducible measurements of fluorescence lifetimes of the macula in healthy subjects. By using a custom-built software, we were able to quantify fluorescence lifetimes within the ETDRS grid. Establishing a clinically accessible standard against which to measure FAF lifetimes within the retina is a prerequisite for future studies in retinal disease.

Fluorescence lifetime imaging of the ocular fundus in mice

PURPOSE Fundus autofluorescence (AF) is characterized not only by its intensity or excitation and emission spectra but also by the lifetimes of the fluorophores. Fluorescence lifetime is influenced by the fluorophores microenvironment and may provide information about the metabolic tissue state. We report quantitative and qualitative autofluorescence lifetime imaging of the ocular fundus in mice. METHODS A fluorescence lifetime imaging ophthalmoscope (FLIO) was used to measure fluorescence lifetimes of endogenous fluorophores in the murine retina. FLIO imaging was performed in 1-month-old C57BL/6, BALB/c, and C3A.Cg-Pde6b(+)Prph2(Rd2)/J mice. Measurements were repeated at monthly intervals over the course of 6 months. For correlation with structural changes, an optical coherence tomogram was acquired. RESULTS Fundus autofluorescence lifetime images were readily obtained in all mice. In the short spectral channel (498-560 nm), mean ± SEM AF lifetimes were 956 ± 15 picoseconds (ps) in C57BL/6; 801 ± 35 ps in BALB/c mice; and 882 ± 37 ps in C3A.Cg-Pde6b(+)Prph2(Rd2)/J mice. In the long spectral channel (560-720 nm), mean ± SEM AF lifetimes were 298 ± 14 ps in C57BL/6 mice, 241 ± 10 ps in BALB/c mice, and 288 ± 8 ps in C3A.Cg-Pde6b(+)Prph2(Rd2)/J mice. There was a general decrease in mean AF lifetimes with age. CONCLUSIONS Although fluorescence lifetime values differ among mouse strains, we found little variance within the groups. Fundus autofluorescence lifetime imaging in mice may provide additional information for understanding retinal disease processes and may facilitate monitoring of therapeutic effects in preclinical studies.

Spectral-domain Optical Coherence Tomography Findings after Severe Exogenous Endophthalmitis

Abstract Purpose: To report outcomes and assess structural changes in the retina in patients with severe endophthalmitis. Methods: Retrospective, nonrandomized, interventional case series at a tertiary referral centre. Spectral domain optical coherence tomography (OCT) images of both eyes were acquired at least 5 months after pars plana vitrectomy. OCT images were analyzed using retinal layer segmentation. Results: Nine patients (46–80 years of age) were included in this study. Average ETDRS visual acuity before treatment was 23 letters and improved to 74 letters. In our cohort we did not find a generalized reduction of retinal layers using automated layer segmentation. Conclusion: Our findings suggest that prompt treatment of severe endophthalmitis with intravitreal antibiotics followed by pars plana vitrectomy may lead to excellent visual outcomes with minimal damage to the retinal architecture.

Fundus Autofluorescence Lifetime Patterns in Retinitis Pigmentosa

Purpose We investigated whether fundus autofluorescence (FAF) lifetimes in patients with retinitis pigmentosa display a disease-specific lifetime pattern. Methods Fundus autofluorescence lifetime imaging ophthalmoscopy (FLIO) was performed in two spectral channels (498-560 and 560-720 nm) after excitation with a 473 nm pulsed laser in patients with retinitis pigmentosa and compared to healthy controls of a similar age range. Corresponding FAF intensity and spectral domain optical coherence tomography (OCT) data, as well as best corrected visual acuity (BCVA) were acquired and compared to fluorescence lifetime data. Results We investigated 43 eyes from 43 patients with retinitis pigmentosa (mean age 45 ± 15 years) and compared them to eyes of 13 age-matched healthy participants. Mean FAF lifetimes were prolonged in areas of photoreceptor atrophy with preserved retinal pigment epithelium (RPE) (P = 0.0036) and even longer in areas with total atrophy of photoreceptors and RPE (P = 0.0002). The prevalence of perifoveal ring structures characterized by prolonged fluorescence lifetimes in FLIO was higher (63% vs. 49%) and the rings were wider compared to the hyperautofluorescent rings in qualitative fundus autofluorescence intensity images. In the central fovea with intact retinal layer structure identified by OCT, fluorescence lifetimes were slightly prolonged compared to those of age-matched healthy controls (short spectral channel [SSC], P = 0.0044; long spectral channel [LSC], P = 0.0128). Short lifetimes within the macular center were negatively correlated with BCVA (R2 = 0.33, P < 0.0001) as well as the greatest diameter of the ellipsoid band in OCT. Conclusions FLIO in retinitis pigmentosa reveals characteristic patterns that allow identification of areas of photoreceptor atrophy, RPE atrophy, and remaining photoreceptor segments in areas of RPE atrophy. Fluorescence lifetimes can be used to identify ellipsoid zone loss that correlates with functional parameters.

Fluorescence Lifetimes of Drusen in Age-Related Macular Degeneration

Purpose The purpose of this study was to characterize fundus autofluorescence lifetimes of retinal drusen in patients with AMD. Methods Patients with AMD and retinal drusen and healthy controls of similar age were examined. A fluorescence lifetime imaging ophthalmoscope was used. Retinal autofluorescence was excited using a 473-nm pulsed laser, and fundus autofluorescence lifetimes of the central retina (30°) were measured in two distinct spectral channels (short: 498 to 560 nm [SSC]; long: 560 to 720 nm [LSC]). Mean retinal autofluorescence lifetimes, corresponding fundus autofluorescence intensity images, spectral domain optical coherence tomography, color fundus images, and clinical data were investigated. Patients were analyzed in two distinct groups (soft drusen and reticular pseudodrusen) and compared with control subjects. Results Sixty-four eyes of 64 patients with AMD and retinal drusen (age: mean ± SD, 78 ± 8.5 years; range, 59 to 94 years) were investigated and compared with a control group of 20 age-matched healthy subjects. Mean retinal autofluorescence lifetimes in patients with AMD was significantly prolonged compared with the healthy control eyes (mean ± SEM: SSC, 486 ± 18 vs. 332 ± 11 ps, P < 0.0001; LSC: 493 ± 9 vs. 382 ± 17 ps, P < 0.0001). Areas of drusen featured a wide range of fluorescence lifetime values. Long lifetimes were identified in areas of atrophy and in areas of intraretinal hyperreflective deposits. Short lifetimes corresponded to deposits within the photoreceptor outer segment band. Conclusions Mean retinal autofluorescence lifetimes in AMD patients are significantly prolonged. Intraretinal deposits cause prolonged lifetimes, whereas deposits in the area of the outer photoreceptor segments lead to short fluorescence lifetimes.

OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY FEATURES OF TORPEDO MACULOPATHY.

BACKGROUND/PURPOSE To investigate the retinal and choroidal vasculature in patients with torpedo maculopathy with optical coherence tomography-angiography (OCT-A). METHODS Retrospective case series of four patients who were examined at the department of Ophthalmology at the University Hospital Bern. Main Outcome was the lesion size over time in OCT-A and fundus autofluorescence. RESULTS Three patients had Type I and 1 patient had Type II torpedo maculopathy. Torpedo maculopathy lesion size remained stable in all patients over a mean period of observation of three years in OCT-A and fundus autofluorescence. The choriocapillaris network was attenuated focally within the lesion in OCT-A in all four cases. The lesion size in fundus autofluorescence was 2.77 mm and therefore comparable with the lesion size in OCT-A of 2.75 mm. CONCLUSION OCT-A signal of the choriocapillaris was reduced within the cleft in both types of torpedo maculopathy. Whether the changes represent the primary site of malformation or whether these findings are the consequence of a defect in the retinal pigment epithelium remains speculative.