研究目的
To investigate fluorescence lifetime characteristics in patients with geographic atrophy (GA) in eyes with age-related macular degeneration and to correlate the measurements with clinical data and optical coherence tomography (OCT) findings.
研究成果
Fluorescence lifetime analysis of areas with GA provides specific lifetime patterns. Short fluorescence lifetimes within the macular center may provide information about the integrity of the outer plexiform layer and outer nuclear layer. The analysis of GA borders by FLIO might emerge as a useful tool for the visualization of local tissue remodeling and disease monitoring.
研究不足
The number of included patients is limited (41 eyes), and larger cohorts are needed to investigate pattern-specific characteristics in different GA subgroups. Additional ex vivo lifetime data and longitudinal follow-up examinations are required to dissect influences and identify markers for disease progression.
1:Experimental Design and Method Selection:
The study used a fluorescence lifetime imaging ophthalmoscope (FLIO) to measure retinal autofluorescence lifetimes in two spectral channels (short: 498–560 nm, long: 560–720 nm). A biexponential decay model was applied for analysis, and data were correlated with best corrected visual acuity (BCVA) and OCT findings.
2:Sample Selection and Data Sources:
41 eyes of 41 patients with GA due to AMD were recruited from the Outpatient Department of Ophthalmology at Bern University Hospital. Exclusion criteria included neovascular AMD, other macular diseases, and significant lens opacities.
3:List of Experimental Equipment and Materials:
Fluorescence lifetime imaging ophthalmoscope (based on Heidelberg Spectralis HRA+OCT system), time-correlated single-photon counting modules with hybrid photon-counting detectors (Becker&Hickl), software (SPCImage
4:6, FLIO reader software), OCT device (Heidelberg Spectralis HRA+OCT), color fundus camera (Zeiss FF 450plus), and pupil dilation agents (tropicamid 5%, phenylephrine hydrochloride 5%). Experimental Procedures and Operational Workflow:
Patients underwent BCVA assessment, pupil dilation, fundus examination, color fundus imaging, FAF imaging, OCT scans, and FLIO imaging. FLIO images were acquired with a 473-nm pulsed laser, and data were analyzed using specified software with a binning factor and biexponential fitting.
5:Data Analysis Methods:
Statistical analysis was performed using Prism Graph Pad and SigmaPlot, including one-way ANOVA, Tukey's posttest, linear regression, and multiple linear regression. Fluorescence lifetime data were analyzed within ETDRS grid areas and correlated with OCT measurements and BCVA.
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software
SPCImage 4.6
Becker&Hickl
Used for analysis of recorded lifetime data, applying a biexponential decay model.
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color fundus camera
Zeiss FF 450plus
Zeiss
Used for obtaining color fundus images.
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fluorescence lifetime imaging ophthalmoscope
based on HRA Spectralis system
Heidelberg Engineering
Used for imaging and measuring retinal autofluorescence lifetimes in patients with geographic atrophy.
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hybrid photon-counting detectors
Becker&Hickl
Used for detecting emitted fluorescence light in the FLIO device.
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FLIO reader software
ARTORG Center for Biomedical Engineering Research, University of Bern
Used for further analysis of fluorescence lifetime data.
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OCT device
Heidelberg Spectralis HRA+OCT
Heidelberg Engineering
Used for high-resolution spectral-domain optical coherence tomography scans of the macula.
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statistical analysis software
Prism 6
GraphPad Software, Inc.
Used for statistical analysis, including one-way ANOVA and Tukey's posttest.
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statistical analysis software
SigmaPlot Version 12.3
Systat Software, Inc.
Used for multiple linear regression analysis.
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