Background: Fundus autofluorescence (AF) is derived from the lipofuscin contained by the retinal pigment epithelial cells. Using a scanning laser ophthalmoscope, two- dimensional AF measurements of the ocular fundus can be achieved. Directly after conventional photocoagulation and also after selective RPE laser treatment (SRT) with ophthalmoscopically non- visi- ble laser lesions, irradiated areas reveal reduced AF, indicating RPE damage. Since the green treatment laser beam could also be used for AF excitation, the aim of this study was to evaluate whether absolute measurements of AF can be performed, and also possible changes in AF detected, online during SRT. Methods: SRT was carried out by use of a frequency- doubled Nd:YLF laser (wavelength 527 nm, pulse duration 1.7 μ s, repetition rate 500 and 100 Hz, number of pulses 100 and 30, single pulse energy 50- 130 μ J) in vitro (porcine RPE; retinal spot size 160 μ m) and during patient treatment (retinal spot size 176 μ m). During irradiation, fluorescence light from the RPE was decoupled from the laser light inside the slit lamp and detected by a photomultiplier or photodiode at wavelengths above 550 nm. Additionally, temperature- dependent fluorescence intensity measurements of A2- E, the main fluorescent component of lipofuscin, were performed in a different in- vitro setup. Results: The intensity of AFdecreased over the number of applied pulses during laser irradiation, and this trend was more pronounced in porcine RPE samples than during human treatment. In vitro, the AF intensity decreased by about 22% ; however, only a weak signal was detected. When treating patients, the AF intensity was strong and the rate of decay of fluorescence intensity with number of pulses was greater when irradiating at 500 Hz than at the 100 Hz repetition rate. However, for both repetition rates the AF decay was merely up to 6- 8% over the number of pulses per laser spot. Fluorescence intensity of A2- E decreased linearly with increasing temperature at about 1% per 1° C and was completely reversible. Conclusions: Online measurements of AF d uring selective RPE laser treatment are possible and reveal a decay in AFas a function of the number of laser pulses applied to the RPE.If A2- E results can be transferred to RPE fluorescence, the AFdecay could be related to the temperature increase within the tissue during treatment. Further clinical studies- in SRT as well as in conventional laser photocoagulation- might be able to show online AF changes on different areas of the retina and on different pathologies. Due to the temperature dependence of the fluorescence, on- line AF measurements during laser treatments such as photocoagulation or TTT may be able to be used as a real- time method for temperature monitoring.
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