Optical microsurgery of the retinal pigment epithelium (RPE) requires reliable real-time dosimetry to prevent unwanted overexposure of the neuroretina. The system used in this experiment implements optical coherence tomography (OCT) to detect the intentional elimination of RPE cells. We evaluated the performance of OCT dosimetry in terms of its ability to detect RPE cell damage caused by microsecond laser pulses of varying duration. Therefore, ex-vivo porcine RPE choroid sclera explants were embedded in an artificial eye and exposed to single laser pulses of 2–20 µs duration (wavelength: 532 nm, exposure area: 120 × 120 µm2, intensity modulation factor: 1.3). Simultaneously, time-resolved OCT M-scans were recorded (central wavelength: 870 nm, scan rate: 33 kHz). Post-irradiation, RPE cell damage was quantified using a calcein-AM viability assay and compared with an OCT-dosimetry algorithm. The results of our experiments show that the OCT-based analysis successfully predicts RPE cell damage. At its optimal operating point, the algorithm achieved a sensitivity of 89% and specificity of 94% for pulses of 6 µs duration and demonstrated the ability to precisely control radiant exposure of a wide range of pulse durations towards selective real-time laser microsurgery.
Selective retina therapy (SRT) is a short pulse (µs-regime) alternative to conventional laser photocoagulation (LPC) for treatment of retinal diseases. LPC leads to collateral damage of retinal layers adjacent to the retinal pigment epithelium (RPE), including healthy, non-regenerative photoreceptors due to the high thermal load, whereas in SRT, RPE cells are destroyed by microbubbles without damaging the neuronal retina. A novel experimental SRT laser operating at 532 nm wavelength can deliver 2 -20 μs pulse sequences. Its tight integration into an upgraded diagnostic SPECTRALIS system combines beam control for treatment planning with real-time optical coherence tomography (OCT) overexposure protection of the photoreceptors. This "Spectralis Centaurus" system, was built and preliminary tested on porcine ex-vivo samples, reaching an unprecedented accuracy with unique planning and follow-up capabilities for upcoming clinical cellular level micro-surgery. The combination of OCT with SRT selectively limits cell death to the RPE by precisely controlling energy deposition while optically monitoring tissue response.
Selective retina therapy (SRT) is currently used in clinical studies to treat several chorioretinal diseases. For SRT a laser pulse duration of 1,7 µs is currently used. At this pulse duration the retinal pigment epithelium (RPE) cells are destroyed by transient microbubbles without damaging the neuronal retina. So far it is unclear whether slightly longer laser pulses are still acting thermomechanically or whether thermal effects show responsible for cell damage close above damage threshold. In order to investigate the damage threshold increase with pulse duration, a novel laser with adjustable pulse duration in the range of 2-20 µs was used to investigate RPE damage on ex-vivo porcine RPE explants. The specimen were fixed in an eye model and were exposed to laser pulse energies ranging from 15-150 µJ with a top hat square of 120x120µm², exhibiting a spatial intensity modulation factor of 1,3. Viability tests using binary evaluation result in threshold values with peak radiant exposures of 233 mJ/cm 2 and 389 mJ/cm 2 for 2 µs and 20 µs laser durations, respectively. An almost logarithmic increase of the threshold radiant exposure over pulse duration was found.
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