Search

Purpose To investigate the most peripheral corneal nerve plexus using high-resolution micro-optical coherence tomography (µOCT) imaging and to assess µOCT's clinical potential as a screening tool for corneal and systemic diseases. Methods An experimental high-resolution (1.5 × 1.5 × 1 µm) µOCT setup was applied for three-dimensional imaging of the subbasal nerve plexus in nonhuman primates (NHPs) and swine within 3 hours postmortem. Morphologic features of subbasal nerves in µOCT were compared to β3 tubulin-stained fluorescence confocal microscopy (FCM). Parameters such as nerve density, nerve distribution, and imaging repeatability were evaluated, using semiautomatic image analysis in form of a custom corneal surface segmentation algorithm and NeuronJ. Results Swine and NHP corneas showed the species-specific nerve morphology in both imaging modalities. Most fibers showed a linear course, forming a highly parallel pattern, converging in a vortex with overall nerve densities varying between 9.51 and 24.24 mm/mm2. The repeatability of nerve density quantification of the µOCT scans as approximately 88% in multiple image recordings of the same cornea. Conclusions Compared to the current gold standard of FCM, µOCT's larger field of view of currently 1 × 1 mm increases the conclusiveness of density measurements, which, coupled with µOCT's feature of not requiring direct contact, shows promise for future clinical application. The nerve density quantification may be relevant for screening for systemic disease (e.g., peripheral neuropathy). Translational Relevance Technological advances in OCT technology may enable a quick assessment of corneal nerve density, which could be valuable evaluating ophthalmic and systemic peripheral innervation.

Background and objectives A recent generation of 5,500 nm wavelength carbon monoxide (CO) lasers could serve as a novel tool for applications in medicine and surgery. At this wavelength, the optical penetration depth is about three times higher than that of the 10,600 nm wavelength carbon dioxide (CO2 ) laser. As the amount of ablation and coagulation is strongly influenced by the wavelength, we anticipated that CO lasers would provide extended coagulation zones, which could be beneficial for several medical applications, such as tissue tightening effects after laser skin resurfacing. Until now, the 1,940 nm wavelength thulium fiber (Tm:fiber) laser is primarily known as a non-ablative laser with an optical penetration depth that is eight times higher than that of the CO2 laser. The advantage of lasers with shorter wavelengths is the ability to create smaller spot sizes, which has a determining influence on the ablation outcome. In this study, the ablation and coagulation characteristics of a novel CO laser and a high power Tm:fiber laser were investigated to evaluate their potential application for fractional ablation of the skin. Study design/materials and methods Laser-tissue exposures were performed using a novel CO laser, a modified, pulse-width-modulated CO2 laser, and a Tm:fiber laser. We used discarded ex vivo human skin obtained from abdominoplasty as tissue samples. Similar exposure parameters, such as spot size (108-120 μm), pulse duration (2 milliseconds), and pulse energy (~10-200 mJ) were adjusted for the different laser systems with comparable temporal pulse structures. Laser effects were quantified by histology. Results At radiant exposures 10-fold higher than the ablation threshold, the CO laser ablation depth was almost two times deeper than that of the CO2 laser. At 40-fold of the ablation threshold, the CO laser ablation was 47% deeper. The ablation craters produced by the CO laser exhibited about two times larger coagulation zones when compared with the CO2 laser. In contrast, the Tm:fiber laser exhibited superficial ablation craters with massive thermal damage. Conclusions The tissue ablation using the Tm:fiber laser was very superficial in contrast to the CO laser and the CO2 laser. However, higher etch depths should be obtainable when the radiant exposure is increased by using higher pulse energies and/or smaller spot sizes. At radiant exposures normalized to the ablation threshold, the CO laser was capable of generating deeper ablation craters with extended coagulation zones compared with the CO2 laser, which is possibly desirable depending on the clinical goal. The effect of deep ablation combined with additional thermal damage on dermal remodeling needs to be further confirmed with in vivo studies. Lasers Surg. Med. © 2020 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Purpose To image, track and map the nerve fiber distribution in excised rabbit corneas over the entire stromal thickness using micro-optical coherence tomography (µOCT) to develop a screening tool for early peripheral neuropathy. Methods Excised rabbit corneas were consecutively imaged by a custom-designed µOCT prototype and a commercial laser scanning fluorescence confocal microscope. The µOCT images with a field of view of approximately 1 × 1 mm were recorded with axial and transverse resolutions of approximately 1 µm and approximately 4 µm, respectively. In the volumetric µOCT image data, network maps of hyper-reflective, branched structures traversing different stromal compartments were segmented using semiautomatic image processing algorithms. Furthermore, the same corneas received βIII-tubulin antibody immunostaining before digital confocal microscopy, and a comparison between µOCT image data and immunohistochemistry analysis was performed to validate the nerval origin of the tracked network structures. Results Semiautomatic tracing of the nerves with a high range of different thicknesses was possible through the whole corneal volumes, creating a skeleton of the traced nerves. There was a good conformity between the hyper-reflective structures in the µOCT data and the stained nerval structures in the immunohistochemistry data. Conclusions This article demonstrates nerval imaging and tracking as well as a spatial correlation between µOCT and a fluorescence corneal nerve standard for larger nerves throughout the full thickness of the cornea ex vivo. Translational relevance Owing to its advantageous properties, µOCT may become useful as a noncontact method for assessing nerval structures in humans to screen for early peripheral neuropathy.

BACKGROUND Traditionally, fractional laser treatments are performed with focused laser sources operating at a fixed wavelength. Using a tunable laser in the mid-infrared wavelength range, wavelength-dependent absorption properties on the ablation process and thermal damage formation were assessed with the goal to obtain customizable tissue ablations to provide guidance in finding optimized laser exposure parameters for clinical applications. METHODS Laser tissue experiments were carried out on full thickness ex vivo human abdominal skin using a mid-infrared tunable chromium-doped zinc selenide/sulfide chalcogenide laser. The laser has two independent channels: a continuous wave (CW) output channel which covers a spectrum ranging from 2.4 μm to 3.0 μm with up to 9.2 W output power, and a pulsed output channel which ranges from 2.35 μm to 2.95 μm. The maximum pulse energy of the pulsed channel goes up to 2.8 mJ at 100 Hz to 1,000 Hz repetition rate with wavelength-dependent pulse durations of 4-7 ns. RESULTS Total ablation depth, ablation efficiency, and coagulation zone thickness were highly correlated to wavelength, pulse width, and pulse energy. Using the same total radiant exposure at 2.85 μm wavelength resulted in 10-times smaller coagulation zones and 5-times deeper ablation craters for one hundred 6 ns pulses compared to one 100 ms pulse. For a fixed pulse duration of 6 ns and a total radiant exposure of 2.25 kJ/cm2 the ablation depth increased with longer wavelengths. CONCLUSION The tunable laser system provides a useful research tool to investigate specific laser parameters such as wavelength on lesion shape, ablation depth and thermal tissue damage. It also allows for customization of the characteristics of laser lesions and therefore facilitates the selection of suitable laser parameters for optimized fractional laser treatments. Lasers Surg. Med. 50:961-972, 2018.© 2018 Wiley Periodicals, Inc.

PURPOSE: To evaluate a new non-ablative and adjustable procedure for laser ablative refractive corneal surgery in hyperopia using the injection of a biocompatible liquid filler material into a stromal pocket. METHODS: A total of 120 stromal pockets were created using a clinical femtosecond laser system in 96 rabbit corneoscleral discs and 24 whole globes. Pockets were cut at a depth of 120 or 250 µm below the epithelial surface. Hyaluronic acid was injected manually into the pocket. To determine the refractive changes, three-dimensional optical coherence tomography images and a specifically developed picture recognition Matlab (The Mathworks) routine were used. RESULTS: After injection, a steepening of the anterior and flattening of the posterior corneal surface was observed, which led to hyperopic correction. The two main factors determining the amount of correction were the pocket depth and the injected volume. After the pocket was homogeneously filled, an initial refractive increase was observed, followed by a linear relation between the injected volume and the refraction increase. CONCLUSIONS: This possible clinical protocol for controlled refraction correction of hyperopia suggests a potential readjustable clinical application. [ J Refract Surg . 2020;36(6):406–414.]

Purpose Photochemical crosslinking of the sclera is an emerging technique that may prevent excessive eye elongation in pathologic myopia by stiffening the scleral tissue. To overcome the challenge of uniform light delivery in an anatomically restricted space, we previously introduced the use of flexible polymer waveguides. We presently demonstrate advanced waveguides that are optimized to deliver light selectively to equatorial sclera in the intact orbit. Methods Our waveguides consist of a polydimethylsiloxane cladding and a polyurethane core, coupled to an optical fiber. A reflective silver coating deposited on the top and side surfaces of the waveguide prevents light leakage to nontarget, periorbital tissue. Postmortem rabbits were used to test the feasibility of in situ equatorial sclera crosslinking. Tensometry measurements were performed on ex vivo rabbit eyes to confirm a biomechanical stiffening effect. Results Metal-coated waveguides enabled efficient light delivery to the entire circumference of the equatorial sclera with minimal light leakage to the periorbital tissues. Blue light was delivered to the intact orbit with a coefficient of variation in intensity of 22%, resulting in a 45 ± 11% bleaching of riboflavin fluorescence. A 2-fold increase in the Young's modulus at 5% strain (increase of 92% P < 0.05, at 25 J/cm2) was achieved for ex vivo crosslinked eyes. Conclusions Flexible polymer waveguides with reflective, biocompatible surfaces are useful for sclera crosslinking to achieve targeted light delivery. We anticipate that our demonstrated procedure will be applicable to sclera crosslinking in live animal models and, potentially, humans in vivo.

Two-Photon Microscopy (TPM) can provide three-dimensional morphological and functional contrast in vivo. Through proper staining, TPM can be utilized to create virtual, H&E equivalent images and thus can improve throughput in histology-based applications. We previously reported on a new light source for TPM that employs a compact and robust fiber-amplified, directly modulated laser. This laser is pulse-to-pulse wavelength switchable between 1064 nm, 1122 nm, and 1186 nm with an adjustable pulse duration from 50ps to 5ns and arbitrary repetition rates up to 1MHz at kW-peak powers. Despite the longer pulse duration, it can achieve similar average signal levels compared to fs-setups by lowering the repetition rate to achieve similar cw and peak power levels. The longer pulses lead to a larger number of photons per pulse, which yields single shot fluorescence lifetime measurements (FLIM) by applying a fast 4 GSamples/s digitizer. In the previous setup, the wavelengths were limited to 1064 nm and longer. Here, we use four wave mixing in a non-linear photonic crystal fiber to expand the wavelength range down to 940 nm. This wavelength is highly suitable for imaging green fluorescent proteins in neurosciences and stains such as acridine orange (AO), eosin yellow (EY) and sulforhodamine 101 (SR101) used for histology applications. In a more compact setup, we also show virtual H&E histological imaging using a direct 1030 nm fiber MOPA.

We demonstrate the highest resolution (1.5×1.5×1 µm) micrometer optical coherence tomography (µOCT) imaging of the morphologic micro-structure of excised swine and non-human primate corneas. Besides epithelial, stromal, and endothelial cell morphology, this report focuses on investigating the most peripheral corneal nerve fibers, the nerve fibers of the subbasal plexus (SBP). Alterations of SBP nerve density and composition are reportedly linked to major neurologic disorders, such as diabetic neuropathy, potentially indicating earliest onsets of denervation. Here, the fine, hyperreflective, epithelial nerve structures located just above Bowman’s membrane, are i) visualized using our µOCT prototype, ii) validated by comparison to fluorescence confocal microscopy (including selective immunohistochemical staining), and iii) segmented using state-of-the-art image processing. Here, we also introduce polarization sensitive (PS) µOCT imaging, demonstrating, to the best of our knowledge, the highest resolution corneal PS-OCT scans reported to date.

Selective retina therapy (SRT) targets the retinal pigment epithelium (RPE) with pulsed laser irradiation by inducing microbubble formation (MBF) at the intracellular melanin granula, which leads to selective cell disruption. The following wound healing process rejuvenates the chorio-retinal junction. Pulse energy thresholds for selective RPE effects vary intra- and interindividually. We present the evaluation of an algorithm that processes backscattered treatment light to detect MBF as an indicator of RPE cell damage since these RPE lesions are invisible during treatment. Eleven patients with central serous chorioretinopathy and four with diabetic macula edema were treated with a SRT system, which uses a wavelength of 527 nm, a repetition rate of 100 Hz, and a pulse duration of 1.7 μs. Fifteen laser pulses with stepwise increasing pulse energy were applied per treatment spot. Overall, 4626 pulses were used for algorithm parameter optimization and testing. Sensitivity and specificity were the metrics maximized through an automatic optimization process. Data were verified by fluorescein angiography. A sensitivity of 1 and a specificity of 0.93 were achieved. The method introduced in this paper can be used for guidance or automatization of microbubble-related treatments like SRT or selective laser trabeculoplasty.

Aims To evaluate selective retina therapy (SRT) as a treatment of acute central serous chorioretinopathy. Methods 30 eyes of 30 patients with central serous chorioretinopathy of at least a 3 months9 duration were recruited. 14 eyes were randomised to an SRT group (Q-switched neodymium-doped yttrium lithium fluoride (Nd:YLF) laser, wavelength 527 nm, t=1.7 μs, energy 100–370 μJ, spot diameter 200 μm, pulse repetition rate 100 Hz,) and 16 eyes to a control group. After 3 months of follow-up, patients in the control group with persistence of subretinal fluid (SRF) were allocated to a cross-over group, treated with SRT and followed up for further 3 months. The main outcome measures were change of best-corrected Early Treatment Diabetic Retinopathy Study visual acuity (BCVA) and SRF. Results At 3 months of follow-up, the mean (SD) improvement of BCVA was significantly greater after SRT than in the control group: 12.7 (7.2) versus 6.3 (8.9) letters (p=0.04). SRF had decreased significantly more after SRT as compared with that the control group: 203 (136) μm versus 41 (150) μm (p=0.005). In eight eyes allocated to the cross-over group, the mean BCVA had increased during 3 months of follow up before SRT by 1.4 (5.2) letters and continued to increase during 3 months following SRT by 7.4 (6.3) letters, while SRF increased by 39.5 (160.2) μm before SRT and decreased by 151.5 (204.9) μm after SRT. In six of the eight eyes, SRF had completely resolved 3 months after SRT. Conclusions SRT appears to expedite functional recovery and the re-absorption of SRF as compared with that in untreated controls. A larger prospective, randomised phase 3 confirmative patient study is warranted. Trial registration number NCT00987077

Background Optical coherence tomography (OCT) is new diagnostic procedure that has rapidly evolved in the last years. The recently developed spectral domain OCT allows one to increase the imaging speed by a hundred times compared to the first generation time domain OCT and enables three-dimensional imaging as well as real-time imaging of fast moving structures. Volumetric imaging improves the quantitative measurement of morphology and the evaluation of temporal changes. In addition, an exact correlation with images acquired with other imaging modalities is possible. Real-time imaging enables also the use of OCT during examinations with the slit-lamp and during ophthalmological surgery. Methods A spectral domain OCT was adapted to a slit lamp. 70 patients (91 eyes) were examined at the anterior or posterior segment of the eye. Images of healthy structures and different pathologies were compared to OCT images obtained with Stratus 3, Spectralis, and the SL-OCT. To demonstrate the feasibility of OCT during surgery, spectral domain OCT devices working with 20,000 and 210,000 A scans per second were coupled by specially developed optics to the camera port of a surgical microscope. The device was tested with phantoms and enucleated pig eyes. Results and conclusions A 5 kHz spectral domain OCT can image the retina during slit lamp-based indirect ophthalmoscopy with a quality similar to that of the Stratus 3. In addition, relevant structures of the anterior segment were imaged. Here, compared to commercially available devices, the imaged field was smaller and the angle of the anterior chamber was not directly visible due to the 830 nm wavelength, which had to be used for retinal imaging. Through the surgical microscope, a volumetric imaging of epithelium, Bowman's, Descemet's membranes, limbus, iris, lens, conjunctiva and sclera was demonstrated with several tens of centimetre working distance. Instruments and incisions in the cornea were visualised with 20 microm precision. Real-time imaging and visualisation of volumetric OCT data were also demonstrated. In principle, all technical problems of an intraoperative use of OCT have been solved and a clinical trial will start in the near future. OCT has the potential to improve the precision of surgical interventions and may even enable new interventions.

The scientific background of laser photocoagulation of the ocular fundus was studied extensively by several investigators in the 1970 s and 1980 s. The basic principles were successfully resolved during that time and clinical consequences for proper application of the laser photocoagulation for various diseases were deduced. The present paper gives an overview about the physical basics of laser-tissue interactions during and after retinal laser treatment and the particular laser strategies in the treatment of different retinal diseases. Thus, it addresses the issue of the impact on tissue of laser parameters as wavelength, spot size, pulse duration and laser power. Additionally, the different biological tissue reactions after laser treatment are presented, such as, e. g., for retinopexia or macular treatments as well as for diabetic retinopathies. Specific laser strategies such as the selective laser treatment of the RPE (SRT) or the transpupillary thermotherapy (TTT) are presented and discussed.

The control of the temperature increase is an important issue in retinal laser treatments. Within the fundus of the eye heat, generated by absorption of light, is transmitted by diffusion in the retinal pigment epithelium and in the choroid and lost by convection due to the choroidal blood flow. The temperature can be spatially and temporally determined by solving the heat equation. In a former analytical model this was achieved by assuming uniform convection for the whole fundus of the eye. A numerical method avoiding this unrealistic assumption by considering convective heat transfer only in the choroid is used here to solve the heat equation. Numerical results are compared with experimental results obtained by using a novel method of noninvasive optoacoustic retinal temperature measurements in rabbits. Assuming global convection the perfusion coefficient was evaluated to 0.07 s � 1 , whereas a value of 0.32 s � 1 – much closer to values found in the literature (between 0.28 and 0.30 s � 1 ) – was obtained when choroidal convection was assumed, showing the advantage of the numerical method. The modelling of retinal laser treatment is thus improved and could be considered in the future to optimize treatments by calculating retinal temperature increases under various tissues and laser properties.

Background and Objectives: The therapeutic effect of laser treatment for macular diseases is related to the damage to the retinal pigment epithelium (RPE) and the subsequent restoration of the defect due to RPE proliferation. In contrast to conventional laser treatment, it is possible to damage the RPE selectively and to spare the photoreceptors by using repetitive microsecond laser pulses. It was the aim of the study to investigate the influence of pulse duration and number of pulses on angiographically and ophthalmoscopically visible retinal damage thresholds in order to optimize treatment modalities. Study Design/Materials and Methods: In total, 625 laser lesions with various parameters were applied to the retina in 11 eyes of 6 Chinchilla breed rabbits using an experimental laser system (Nd:YLF at 527 nm). Pulse duration (1.7 microseconds and 200 nanoseconds) and number of pulses (100, 10, and 1 pulses) were varied at a constant repetition rate of 100 Hz. Damage thresholds were determined in terms of ophthalmoscopic and fluorescein angiographic visibility, and the therapeutic window (TW; angiographic ED50 vs. ophthalmoscopic ED50) as well as the safety range (SR; angiographic ED84 vs. ophthalmoscopic ED16) between both thresholds were calculated. Selected laser lesions were evaluated by histology. Results: Generally, the ED50 radiant exposure for angiographic visibility decreases for shorter laser pulses and with an increase in the number of pulses. The TW for both pulse durations (1.7 microseconds and 200 nanoseconds) was wider with 100 pulses than with single pulses. The widest TW was found for 100 pulses at 200 nanoseconds pulse duration (5.9-fold above the angiographic threshold), and the smallest TW with a factor of 1.6 was found for 1.7 microseconds single pulses. In terms of SR, only irradiation with 100 pulses at 200 nanoseconds pulse duration was associated with a ratio > 2. Independently of pulse duration, histological examination of laser sites 1 hour after irradiation revealed widely intact photoreceptors, while the underlying RPE was damaged. Conclusions: Pulse duration and number of pulses have a significant influence on RPE damage thresholds and consecutively on TW and SR. Because fundus pigmentation in humans may vary intra- and interindividually by a factor of 2, a large TW and ideally also a large SR should be ensured in a clinical treatment context. In rabbits, the safety range with 200 nanoseconds pulses is higher than with the pulse duration of 1.7 microseconds currently in clinical use. These findings suggest the need for clinical pilot studies to prove whether these results can be transposed to the situation in humans. Lasers Surg. Med. 34:206–215, 2004. 2004 Wiley-Liss, Inc.

Background and Objectives Selective treatment of the retinal pigment epithelium (RPE) by repetitively applying green μs-laser pulses is a new method for retinal diseases associated with a degradation of the RPE, which spares the neural retina. We investigated an alternative approach to realize repetitive μs-laser exposure by rapidly scanning a continuous wave (CW)-laser beam across the RPE. Study Design/Materials and Methods An Ar+ laser beam (514 nm) with a diameter of 18.75 μm was repetitively scanned across porcine RPE samples in vitro providing an irradiation time of 1.6 μs per point on the central scan axis. RPE cell damage was investigated by means of the fluorescence viability assay Calcein-AM. Results The ED50 cell damage is 305 mJ/cm2 when applying 10 scans with a repetition rate of 500 Hz. The threshold decreases with the number of scans, a saturation was found at 135 mJ/cm2 with more than 500 exposures applied. The depth of focus in beam direction is 350 μm, defined by an increase of the threshold radiant exposure by 20%. Conclusions Targeting of pigmented cells with high local resolution has been proved with a laser-scanning device. Looking ahead selective RPE-treatment, the adaptation of a laser-scanning device on a slit-lamp or into a modified retina angiograph seems to be an attractive alternative to the pulsed μs laser device. Lasers Surg. Med. 32:252–264, 2003. © 2003 Wiley-Liss, Inc.

* BACKGROUND AND OBJECTIVE: In both clinical and animal studies, it has been shown that repetitive short laser pulses can cause selective retinal pigment epithelium damage (RPE) with sparing of photoreceptors. Our purpose was to determine the ophthalmoscopic and angiographic damage thresholds as a function of pulse durations by using different pulsed laser systems to optimize treatment modalities. * MATERIALS AND METHODS: Chinchilla breed rabbits were narcotized and placed in a special holding system. Laser lesions were applied using a commercial laser slit lamp, contact lens, and irradiation with a frequency-doubled Nd: YLF laser (wavelength: 527 nm; repetition rate: 500 Hz; number of pulses: 100; pulse duration: 5 µß, 1.7 µß, 200 ns) and an argon-ion laser (514 nm, 500 Hz, 100 pulses, 5 µß and 200 ms). In all eyes, spots with different energies were placed into the regio macularis with a diameter of 102 µp? (tophat profile). After treatment, fundus photography and fluorescein angiography were performed and radiant exposure for ED50 damage determined. Speckle measurements at the fiber tips were performed to determine intensity peaks in the beam profile. * RESULTS: Using the Nd:YLF laser system, the ophthalmoscopic ED50 threshold energies were 25.4 µ] (5 µß), 32 µ] (1.7 µß), and 30 µ] (200 ns). The angiographic ED50 thresholds were 13.4 µ] (5 µ8), 9.2 µ] (1.7 µß), and 6.7 µ] (200 ns). With the argon laser, the angiographic threshold for 5 µß pulses was 5.5 µ). The ophthalmoscopic threshold could not he determined because of a lack of power; however, it was >12 µ}. For 200 ms, the ED50 radiant exposures were 20.4 mW ophthalmoscopically and 19.2 mW angiographically. Speckle factors were found to be 1.225 for the Nd: YLF and 3.180 for the argon laser. Thus, the maximal ED50-threshold radiant exposures for the Nd:YLF were calculated to be 362 mj/cm2 (5 µß), 478 mj/cm2 (1.7 µ-s), and 438 mj/cm2 (200 ns) ophthalmoscopically. Angiographically, the thresholds were 189 mj/cm2 (5 µß), 143 mj/cm2 (1.7 µ-s), and 97 mj/cm2 (200 ns). For the argon laser, the maximal ED50 radiant exposure threshold was 170 mj/cm2 angiographically. * CONCLUSION: The gap between the angiographic and the ophthalmoscopic thresholds for the 200 ns regime (4.5 times above angiographic ED50) was wider than for the 1.7 µ$ regime (3.3 times above the angiographic ED50). This would suggest the appropriate treatment would be 200 ns pulses. However, histologies have yet to prove that nonvisible mechanical effects increase with shorter pulse durations and could reduce the "therapeutic window." When comparing the thresholds with 5 µß pulses from the argon and Nd:YLF laser, it demonstrates that intensity modulations in the beam profile must be considered. [Ophthalmic Surg Lasers 200233:400-409]

Laser photocoagulation has been a treatment method for retinal diseases for decades. Recently, studies have demonstrated therapeutic benefits for subvisible effects. A treatment mode based on an automatic feedback algorithm to reliably generate subvisible and visible irradiations within a constant irradiation time is introduced. The method uses a site-individual adaptation of the laser power by monitoring the retinal temperature rise during the treatment using optoacoustics. This provides feedback to adjust the therapy laser power during the irradiation. The technique was demonstrated on rabbits in vivo using a 532-nm continuous wave Nd:YAG laser. The temperature measurement was performed with 523-nm Q-switched Nd:YLF laser pulses with 75-ns pulse duration at 1-kHz repetition rate. The beam diameter on the fundus was 200 μm for both lasers, respectively. The aim temperatures ranged from 50°C to 75°C in 11 eyes of 7 rabbits. The results showed ophthalmoscopically invisible effects below 55°C with therapy laser powers over a wide range. The standard deviation for the measured temperatures ranged from 2.1°C for an aim temperature of 50°C to 4.7°C for 75°C. The ED50 temperature value for ophthalmoscopically visible lesions in rabbits was determined as 65.3°C. The introduced method can be used for retinal irradiations with adjustable temperature elevations.

Summary Irradiation of nanoabsorbers with pico- and nanosecond laser pulses could result in thermal effects with a spatial confinement of less than 50 nm. Therefore absorbing nanoparticles may be used for creating controlled intracellular effects. Conjugates of colloidal gold and alkaline phosphatase (aP) are presented as a model system for investigating protein inactivation in the vicinity of strongly absorbing nanoparticles. aP was coupled either directly or via antibodies to 15 nm gold particles. These conjugates were irradiated with picosecond pulses emitted by a frequency doubled Nd:YLF laser. Denaturation was detected as a loss of protein function with the help of a fluorescence assay. Irradiation with 10 4 pulses resulted in the inactivation of aP at an irradiance of 50 mJ/cm 2 per pulse when the protein was coupled directly to the particles. With similar irradiation parameters a significant inactivation of aP which was coupled not directly but via two antibodies to the gold, was not observed. This shows that inactivating proteins with nanoabsorbers under irradiation with picosecond laser pulses is possible with a high spatial confinement. However, from these experiments it is difficult to determine whether the aP was destroyed thermally or whether it was inactivated by a photochemical reaction. Therefore, further experiments are needed to elucidate the damage mechanisms.

Keywords: Photomedicine group Reference LPAS-ARTICLE-2001-009 Record created on 2007-07-20, modified on 2016-08-08

PURPOSE: To investigate the representation of the corneal structure with optical coherence tomography before and immediately after excimer laser photorefractive keratectomy. METHODS: Twenty-four eyes of 24 patients with myopia and myopic astigmatism were prospectively studied. The corneal thickness and the corneal profile were assessed with slit-lamp–adapted optical coherence tomography preoperatively and immediately after excimer laser photorefractive keratectomy. RESULTS: The attempted mean spherical equivalent of the refractive corrections was −6.7 ± 3.6 (mean ± SD) diopters with a mean calculated stromal ablation depth of 91 ± 38 μm. The corneal optical coherence tomography was reproducible in all patients, demonstrating a mean decrease of central corneal thickness after epithelial debridement and excimer laser photorefractive keratectomy of 118 ± 45 μm. The comparison of the calculated stromal ablation depth and the corneal thickness changes determined by corneal optical coherence tomography revealed a significant linear relationship with a correlation coefficient of 0.88 (P < .001). The flattening of the corneal curvature was confirmed in all patients with the optical coherence tomography system and correlated with the attempted refractive correction (r = .82, P < .001). CONCLUSIONS: The slit-lamp–adapted optical coherence tomography system presented in this study allowed noncontact, cross-sectional, and high-resolution imaging of the corneal configuration. This initial clinical evaluation demonstrated that corneal optical coherence tomography could be a promising diagnostic modality to monitor corneal changes of thickness and curvature before and after excimer laser photorefractive keratectomy.

Three-dimensional topography of perfused vascular structures is possible via confocal laser scanning of intravascular fluorescence. The lateral resolution is given by the spot size of the scanning laser beam (optimally 10 microm at the retina). The axial resolution, however, depends on the accuracy of detection of the surface of the fluorescent structure, which is typically one order of magnitude higher (30 microm at the retina) than the confocal resolution. The vascular structure is stained with an appropriate fluorescent dye prior to the investigation using standard systemic dye injection. Confocal scanning of the fluorescence in planes of different depths within the vascular structure under investigation leads to a three-dimensional data set. Signal processing includes passive eye tracking, lateral averaging and axial determination of the surface of the fluorescent structure. The potential of this new technique is demonstrated by showing the topography of physiological vessel structures as well as of selected vascular diseases such as cone dystrophy, RPE detachment, choroidal haemangioma and retinal laser coagulation. Confocal laser angioscopic fluorescence topography (CLAFT) measures the three-dimensional surface structure of functional (perfused) vasculature and surrounding leakage. CLAFT may help to diagnose and quantify status and time course of vascular diseases.

Purpose To investigate thermomechanical aspects of corneal collagen denaturation as a function of temperature and time and the effect of the induced forces on refractive changes with laser thermokeratoplasty (LTK). Setting Medical Laser Center Lubeck, Lubeck, Germany. Methods In a material-test setup, porcine corneal strips were denatured in paraffin oil at various constant temperatures for 10 and 500 seconds, and the temporal course of the contractive forces was studied under isometric conditions. Typical LTK lesions were performed in porcine eyes in vitro with a continuous-wave infrared laser diode at a wavelength of 1.87 μm for 10 and 60 seconds. The laser power was chosen to achieve comparable denatured volumes at both irradiation times. The refractive changes were measured and analyzed by histologic evaluations and temperature calculations. Results The time course of the induced forces was characterized by a maximal force, which increased almost linearly with temperature, and a residual lower force. After 500 seconds of heating, the highest force was achieved with a temperature of 75°C. With a limited heating period of only 10 seconds, the forces steadily increased with temperature over the entire observation period. Laser thermokeratoplasty produced less refractive change after 10 seconds of irradiation than after 60 seconds, although the laser power was 25% higher in the short heating period. Polarization light microscopy of LTK lesions revealed different stages of thermal damage. Conclusion The course of the contractive forces during and after heating is a complicated function of the spatial time/temperature profile. Laser thermokeratoplasty lesions produced with 2 irradiation times showed different stages of denaturation and induced refractive change.

Background and Objective: Selective photodamage of the retinal pigment epithelium (RPE) is a new technique to treat a variety of retinal diseases without causing adverse effects to surrounding tissues such as the neural retina including the photoreceptors and the choroid. In this study, the mechanism of cell damage after laser irradiation was investigated. Study Design/Materials and Methods: Single porcine RPE-melanosomes and RPE cells were irradiated with a Nd:YLF laser (wavelength l = 527 nm, adjustable pulse duration t = 250 nsec-3 msec) and a Nd:YAG laser (l = 532 nm, t = 8 nsec). Fast flash photography was applied to observe vaporization at melanosomes in suspension. A fluorescence viability assay was used to probe the cells vitality. Results: The threshold radiant exposures for vaporization around individual melanosomes and for ED50 cell damage are similar at 8-nsec pulse duration. Both thresholds increase with pulse duration; however, the ED50 cell damage radiant exposure is 40% lower at 3msec. Temperature calculations to model the onset of vaporization around the melanosomes are in good agreement with the experimental results when assuming a surface temperature of 150°C to initiate vaporization and a homogeneous melanosome absorption coefficient of 8,000 cm ˛1 . Increasing the number of pulses delivered to RPE cells at a repetition rate of 500 Hz, the ED50 value decreases for all pulse durations. However, the behavior does not obey scaling laws such as the N 1/4 equation. Conclusion: The origin of RPE cell damage for single pulse irradiation up to pulse durations of 3 msec can be described by a damage mechanism in which microbubbles around the melanosomes cause a rupture of the cell structure. The threshold radiant exposure for RPE damage decreases with increasing number of pulses applied. Lasers Surg. Med. 27:451‐464, 2000. © 2000 Wiley-Liss, Inc.

Purpose: Pulsed holmium lasers are currently used to correct hyperopia by means of laser thermokeratoplasty (LTK). Series of μs laser pulses are applied with a high repetition rate to induce shrinkage of corneal collagen fibers. The pulsed energy application results in intrastromal temperature peaks of up to 200 °C. A continuously emitting laser diode can – as we demonstrated recently in an invivo study on minipigs – be used for LTK and may be of advantage because the temperature rise is more steady. The aim of this study was to examine the safety, amount, and stability of hyperopic correction of diode LTK on blind human eyes. Methods: We used a laserdiode that was set to continuously emit light at λ = 1.854 μm/μa = 1.04 mm–1(group I, n = 4) or 1.87 μm/μa = 1.92 mm–1 (group II, n = 4). Radiation energy was 100 to 150 mW for 10 s per coagulation. Eight coagulations on a single ring (group I) and 16 coagulations on a double ring (group II) diameter were applied in the cornea concentric to the entrance pupil by means of a vacuum-fixed application mask (group I = conjunctival fixation; group II = corneal fixation) and a handpiece with a focusing optic. Preoperatively as well as 1 week, 1, 2, 3, 6 12 and 18 months postoperative ophthalmologic controls were performed and the corneal refractive power was measured. Results: In group I initial refractive changes of up to + 4.9 D were achieved (1 week postoperative). However, due to the great penetration depth of the laser irradiation, large endothelial defects resulted beneath the stromal coagulations. In group II an initial refractive change of up to + 6.8 D was achieved and as a result of the reduced penetration depth, the endothelial cell damage was much reduced. Partial regression of the refractive effect occured in all subjects, which continued in higher refractive changes during the 2nd postoperative year. The refractive effect at 12 months was + 0.6 to + 1.5 D in group I and + 0.9 to + 5.7 D in group II. At 12 months the induced astigmatism was 0.5 to 2.2 D in group I and 0.3 to 1.6 D in group II. No serious adverse effects were noticed. Conclusion: A continously emitting laser diode working at a wavelength of 1.87 μm can be used to correct hyperopia by means of LTK safely and effectively. Regression occurs predominantly in the first 6 postoperative months. Further studies must be conducted to determine the importance of patient inherent parameters such as age in establishing a nomogram.

During optical breakdown, the energy delivered to the sample is either transmitted, reflected, scattered, or ab- sorbed. Pathways for the division of the absorbed energy are the evaporation of the focal volume, the plasma radiation, and the mechanical effects such as shock wave emission and cav- itation. The partition of laser energy between these channels during breakdown in water was investigated for four selected laser parameters typical for intraocular microsurgery ( 6-ns pulses of 1 and 10 mJ focused at an angle of 22 ,a nd30-ps pulses of 50 mJ and 1m Jfocused at 14 ,a ll at1064 nm). Scattering and reflection were found to be small compared to transmission and absorption during optical breakdown. The ratio of the shock wave energy and cavitation bubble energy was approximately constant (between 1.5:1 and 2:1). These results allowed us to perform a more comprehensive study of the influence of pulse duration ( 100 fs- 76 ns )a nd focus- ing angle (4- 32) on the energy partition by measuring only the plasma transmission and the cavitation bubble energy. The bubble energy was used as an indicator for the total amount of mechanical energy. We found that the absorption at the breakdown site first decreases strongly with decreasing pulse duration, but increases again for < 3p s. The conversion of the absorbed energy into mechanical energy is 90% with ns pulses at large focusing angles. It decreases both with de- creasing focusing angle and pulse duration (to < 15 %f or fs pulses). The disruptive character of plasma-mediated laser effects is therefore strongly reduced when ultrashort laser pulses are used.

The precision of laser-induced effects is often limited by thermal and thermomechanical collateral damage. Adjusting the pulsewidth of the laser to the size of the absorbing structure can at least avoid thermal side effects and facilitates a selective treatment of vessels or pigmented cells. Further extending the precision of thermal effects below cellular dimensions by using nanometer sized particles could open up new fields of applications for lasers in medicine and biology. Calculations show that under irradiation with nano- or picosecond laser pulses gold particles of submicrometer size can easily be heated by several hundred K. High temperatures have to be used for subcellular thermal effects, because heat confinement to such small structures requires the thermal damage to occur in extremely short times. Estimating the denaturation temperature by extrapolating the Arrhenius equation from a time range of minutes and seconds into a time range of nano- and picoseconds leads to temperatures between 370 K-470 K. There is evidence that in aqueous media, due to the surface tension, these temperatures can be generated at the surface of nanometer sized particles without vaporization of the surrounding water. In order to show whether or not an extrapolation of the damage rates over six to nine orders of magnitude gives correct data, a temperature-jump experiment was designed and tested which allows to measure denaturation rates of proteins in the millisecond time range. Denaturation of chymotrypsin was observed within 300 ps at temperatures below 380 K. The rate constants for the unfolding of chymotrypsin followed the Arrhenius equation up to rates of 3000 s/sup -1/.

Endocardial laser revascularization (ELR) is a new technique to treat coronary heart disease in a percutaneous, minimally invasive approach. A holmium laser (/spl lambda/=2.12 /spl mu/m) was developed to emit pulse energies of up to 30 J in order to ablate the desired channels in a single laser pulse. The energy was transmitted by multimode flexible optical waveguides as required for ELR. Ablation dynamics were investigated in two model systems, water serving as blood model and polyacrylamide (PAA) as a transparent tissue phantom. Measurements were undertaken using pulse energies of 12 J at pulse durations of 2.2 and 8 ms with a beam diameter of 1 mm. For comparison with the clinically established method of transmyocardial revascularization (TMR), ablations were also investigated with a standard 800 W TMR CO/sub 2/ laser. The dynamics were recorded with a drum camera and stroboscope illumination providing a high framing rate of a single ablation process. Tissue ablation was quantified with the holmium laser in vitro on porcine heart tissue using pulse energies of up to 20 J. Tissue morphology was evaluated using polarization light microscopy to determine thermal and mechanical collateral damage zones. Oscillating vapor bubble channels were found in water and PAA with all laser systems and parameters used. Quasi-static vapor bubbles are observed in water in the millisecond time range using the holmium laser. CO/sub 2/ laser radiation performed deeper channels in PAA than holmium laser pulses using the same radiant exposure. Channel depths of up to 10 mm were achieved with the holmium laser in myocardial tissue with pulse energies of 17 J, Thermal damage zones of about 150 /spl mu/m for the CO/sub 2/ and 500 /spl mu/m for the holmium laser were found. The orientation of myocardial fibrils significantly influences the shape of the ablated cavities and the thermo-mechanical collateral damage zones. In conclusion, the results are very encouraging and demonstrate the potential of a catheter-based minimal invasive procedure for heart reperfusion using single high energy laser pulses.

Purpose: To investigate suitable application parameters for efficient hyperopic correction by laser thermokeratoplasty (LTK) using mid-infrared laser diodes. Setting: Medical Laser Center Lubeck, Lubeck, Germany. Method: A tunable continuous-wave laser diode in the spectral range between 1.845 and 1.871 pm was used. Transmitted by waveguides, the laser energy was used to induce coagulations on freshly enucleated porcine eyes to increase corneal curvature. The coagulations were equidistantly applied by a fiber-cornea contact and a noncontact focusing device that were adjusted on a ring concentric to the corneal apex. Different laser parameters and application geometries were evaluated. Refractive changes were measured by computer-assisted corneal topography before and after treatment. Polarization light microscopy and temperature calculations were used to analyze the coagulations. Results: Because of the tunability of the laser diode, the influence of the corneal absorption coefficient (between 0.9 and 1.6 mm -1 ) on the refractive change could be measured. A laser power between 125 and 200 mW was adequate to achieve refractive changes up to 10.0 diopters. In the preferable focusing device, the refractive change increased almost logarithmically with the irradiation time up to 15 seconds. The number of coagulations on a fixed application ring showed no significant influence on refractive change; however, it showed an almost linear decrease with increasing ring diameter from 5.0 to 10.0 mm. Histological analysis revealed 3 stages of thermal damage. Conclusion: Diode LTK provided defined and uniform coagulations when using a well-adapted focusing device, resulting in sufficient refractive change. The results indicate that diode LTK is superior to pulsed holmium LTK.

Background and Objective Retinal laser photocoagulation is generally performed by laser pulses of a few hundred milliseconds. The tissue interaction mechanism is a pure thermal interaction mechanism. As pulse duration gets shorter, different, non-thermal interaction mechanisms start to appear. The time domain for a change of tissue interaction mechanism seems to be in the ns and μs range. The goal of this study was to characterize the tissue interaction mechanism with 200-ns laser pulses, which approximate the thermal relaxation time of single melanin granules. Materials and Methods The retinas of 19 eyes of 10 rabbits were irradiated by 10 and 500 repetitive laser pulses (wavelength, 532 nm; repetition rate, 500 Hz; pulse duration, 200 ns; per pulse energy, 0–120 μJ; retinal spot size, 100 μm). The effects were evaluated by fluorescein angiography, ophthalmoscopy and by theoretical thermal calculations. Light microscopy and transmission electron microscopy were additionally performed on lesions irradiated by 500 pulses. Results Single pulse threshold energies for angiographic visibility were 3.5 μJ (10 pulses) and 2.1 μJ (500 pulses), for ophthalmoscopic visibility 9.0 μJ (10 pulses) vs. 8.6 μJ (500 pulses). At energy levels above ophthalmoscopic visibility macroscopically visible bubble formation inside the retina could be observed. This occurred at energy levels of 35 μJ (10 pulses) vs. 17 μJ (500 pulses). Microscopic evaluation of lesions irradiated with 500 pulses and energies at the angiographic threshold showed a damage primarily to the RPE. Additional outer segment damage of the photoreceptors could be found. A gap between damaged RPE cells and the outer segments could be repeatedly found as well as damaged RPE cells, which were detached from intact Bruch's membrane. Temperature calculation shows that temperatures above 100°C may exist around single melanin granules. Conclusion The studies suggest that RPE damage may occur by bubble formation around single melanin granules. Lasers Surg. Med. 22:240–248, 1998. © 1998 Wiley-Liss, Inc.

TARGET The method of optical coherence tomography (OCT) was investigated regarding its suitability and limits for measuring the cornea and the anterior segment of the eye. Furthermore, the stromal expansion of thermally induced lesions in the cornea directly after irradiation was determined within the scope of the laser thermokeratoplasty (LTK). MATERIAL AND METHODS With the experimental scanning OCT system, x-z sections of the anterior eye segment were made with an optical resolution of about 20 microns axially and 25 microns laterally. Freshly enucleated, tonicized porcine eyes were used as model eyes. Thermal lesions were applied with a continuously emitting laser diode (lambda = 1.86 microns) and various radiation parameters. Before and after coagulation, the cornea was viewed from limbus to limbus in a central OCT scan and the individual coagulation source was measured. RESULTS Global and local changes of the thickness of the cornea as well as the distance between cornea and lens were measured with high precision. Thermal lesions in their expansion can be dearly presented and matching well with the histologically stained sections, but are not as exactly defined at the edges due to the limited optical resolution, as known from histological preparations. CONCLUSION With the OCT method quantitative measuring of the anterior eye segment can be performed in vitro and with reduced resolutions also in vivo. Due to the qualitatively good correspondence regarding the dimensions of thermal damage of the cornea with histologically obtained morphometric results, this method can be used for supervision of coagulation directly after LTK as well as for examination of the individual healing process.

Purpose: The aim of this study was to understand the dynamics of ablation products during excimer laser photorefractive keratectomy, and their influence on the formation of central islands. Method: Laser flash photography was used to investigate the dynamics of ablation products during photorefractive keratectomy. The ablation plume over polymethyl methacrylate (PMMA) and porcine cornea targets ablated with different zone diameters was imaged at various times between 10 μsec and 100 msec after the ablating laser pulse. On PMMA targets, the profiles of the resulting ablation craters were measured. Results: In all cases, the ablation products formed a ring vortex. The plume velocities on cornea were found to be approximately twice as fast as on PMMA for all zone diameters. For both materials, the ablation plume evolves faster for smaller zone diameters. Central islands were observed for zone diameters between 1 and 7 mm on PMMA substrates. The islands became more pronounced with increasing zone diameter. Conclusion: A major cause for the formation of central islands was found to be particle redeposition at the center of the ablation zone. Because of the vortex dynamics, redeposition of particles favorably occurs at the center of the ablation zone. Additionally, the dynamics of the ablation plume lead to a concentration of airborne particles over the center of the ablation zone, which also may contribute to the creation of central islands by partial absorption of the next excimer laser pulse.

To reduce unwanted collateral thermal damage to surrounding tissue and organs during laparoscopic laser dissection (cw, wavelength: 1.9μm) of porcine liver water spray was used. Size and amount of the produced water droplets of the water spray were photographed by short time imaging and analyzed by imaging software. At in vivo measurements on fresh porcine liver the depth of thermal damage was reduced by 85 % with water spray and the lateral size of thermal damage at the tissue surface could be reduced by 67%. This results show that especially for laparoscopic laser surgery water spray application might be a useful tool to avoid unwanted collateral thermal damage.

Retinal photocoagulation is an established treatment for various retinal diseases. The temperature development during a treatment can be monitored by applying short laser pulses in addition to the treatment laser light. The laser pulses induce thermoelastic pressure waves that can be detected at the cornea. We present a numerical model to examine the temperature development during the treatment as well as the formation and propagation of the ultrasonic waves. Using the model, it is possible to determine the peak temperature during retinal photocoagulation from the measured signal, and investigate the behaviour of the temperature profile and the accuracy of the temperature determination under varying conditions such as inhomogeneous pigmentation or change in irradiation parameters. It was shown that there is an uncertainty of 2.5‒9% in the determination of the peak temperature when the absorption coefficient between the absorbing layers is varied by a factor of 2. Furthermore the model was extended in order to incorporate the photoacoustic pressure generation and wave propagation. It was shown that for an irradiation pulse duration of 75 ns the resulting pressure wave energy is attenuated by 76 % due to frequency dependent attenuation in water.

Retinal photocoagulation lacks objective dosage in clinical use, thus the commonly applied lesions are too deep and strong, associated with pain reception and the risk of visual field defects and induction of choroidal neovascularisations. Optoacoustics allows real-time non-invasive temperature measurement in the fundus during photocoagulation by applying short probe laser pulses additionally to the treatment radiation, which excite the emission of ultrasonic waves. Due to the temperature dependence of the Gruneisen parameter, the amplitudes of the ultrasonic waves can be used to derive the temperature of the absorbing tissue. By measuring the temperatures in real-time and automatically controlling the irradiation by feedback to the treatment laser, the strength of the lesions can be defined. Different characteristic functions for the time and temperature dependent lesion sizes were used as rating curves for the treatment laser, stopping the irradiation automatically after a desired lesion size is achieved. The automatically produced lesion sizes are widely independent of the adjusted treatment laser power and individual absorption. This study was performed on anaesthetized rabbits and is a step towards a clinical trial with automatically controlled photocoagulation.

We present optical coherence-gated tomography (OCT) in turbid biological tissues. The fast OCT system that is used in this study is a single-mode fiber-optic interferometer with low-coherence light at 830 nm which can perform a cross-sectional image of 250-600 pixels in a few seconds. Preliminary results suggest that OCT can provide high-resolution imaging in low-scattering and high-scattering superficial tissues. In vitro imaging of porcine cornea after being coagulated with a laser shows that OCT is a promising tool for the evaluation of pathological structures in low-scattering tissue. This technique can also be used to diagnose disease in high-scattering tissues like bladder and living skin. In addition, 50 MHz ultrasound images and histological pictures are presented for comparison with OCT.

Flashlamp pumped mid-IR laser systems emitting in the 2-3-/spl mu/m wavelength range are widely used for various medical applications, especially for tissue ablation. Explosive evaporation is inevitably associated with this process due to the short pulse durations of these laser systems and the high absorption of tissue and water in this spectral regime. Tissue displacement and dissection occur in liquid environment as a consequence of the induced cavitation. Depending on the application these processes might enhance the tissue ablation but can also cause adverse tissue effects. The ablation dynamics were investigated by evaluating the change in reflected probe-light intensity re-emitted from the application fiber tip. The ablated cavity and the signal was correlated to fast-flash photographs of the event. Based on this reflection signal a water/tissue discrimination system is introduced which can widely support medical laser applications. In laser sclerostomy ab externo, for example, this approach can be used as a feedback system to automatically control the ablation process. With such a system, adverse effects to adjacent tissue in the anterior chamber of the eye can be minimized.

Laser photocoagulation is an established treatment for a variety of retinal diseases. However, when using the same irradiation parameter, the size and strength of the lesions are unpredictable due to unknown inter- and intraindividual optical properties of the fundus layers. The aim of this work is to investigate a feedback system to generate desired lesions of preselectable strengths by automatically controlling the irradiation time. Optoacoustics were used for retinal temperature monitoring. A 532-nm continuous wave Nd:YAG laser was used for photocoagulation. A 75-ns/523-nm Q-switched Nd:YLF laser simultaneously excited temperature-dependent pressure transients, which were detected at the cornea by an ultrasonic transducer embedded in a contact lens. The temperature data were analyzed during the irradiation by a LabVIEW routine. The treatment laser was switched off automatically when the required lesion strength was achieved. Five different feedback control algorithms for different lesion sizes were developed and tested on rabbits in vivo. With a laser spot diameter of 133???m, five different lesion types with ophthalmoscopically visible diameters ranging mostly between 100 and 200???m, and different appearances were achieved by automatic exposure time control. The automatically controlled lesions were widely independent of the treatment laser power and the retinal pigmentation.

Laser coagulation is a treatment method for many retinal diseases. Due to variations in fundus pigmenta- tion and light scattering inside the eye globe, different lesion strengths are often achieved. The aim of this work is to realize an automatic feedback algorithm to generate desired lesion strengths by controlling the retinal temperature increase with the irradiation time. Optoacoustics afford non-invasive retinal temperature monitoring during laser treatment. A 75 ns∕523 nm Q-switched Nd:YLF laser was used to excite the temperature-dependent pressure amplitudes, which were detected at the cornea by an ultrasonic transducer embedded in a contact lens. A 532 nm continuous wave Nd:YAG laser served for photocoagulation. The ED50 temperatures, for which the prob- ability of ophthalmoscopically visible lesions after one hour in vivo in rabbits was 50%, varied from 63°C for 20 ms to 49°C for 400 ms. Arrhenius parameters were extracted as ΔE ¼ 273 Jmol −1 and A ¼ 3 · 10 44 s −1 . Control algo- rithms for mild and strong lesions were developed, which led to average lesion diameters of 162 � 34 μm and 189 � 34 μm, respectively. It could be demonstrated that the sizes of the automatically controlled lesions were widely independent of the treatment laser power and the retinal pigmentation. © 2012 Society of Photo-Optical Instrumentation

We conducted a study to correlate the retinal temperature rise during photocoagulation to the afterward detected tissue effect in optical coherence tomography (OCT). 504 photocoagulation lesions were examined in 20 patients. The retinal temperature increase was determined in real-time during treatment based on thermoelastic tissue expansion which was probed by repetitively applied ns laser pulses. The tissue effect was examined on fundus images and OCT images of individualized lesions. We discerned seven characteristic morphological OCT lesion classes. Their validity was confirmed by increasing visibility and diameters. Mean peak temperatures at the end of irradiation ranged from approx. 60 °C to beyond 100 °C, depending on burn intensity.

Phase sensitive OCT enables the measurement of thermal expansion in laser irradiated material at high lateral and temporal resolution. In principle, a calculation of the 3D temperature distribution and its temporal evolution should be possible by evaluating the local expansion. This could be utilized for a non-invasive and very fast temperature measurement, e.g. to realize an online dosimetry for photocoagulation. The possibilities of quantitative investigations at high axial and lateral resolution are demonstrated by imaging the reversible thermal expansion in laser irradiated multilayer silicone phantoms.

In retinal laser photocoagulation, constant exposure parameters do not result in identical lesions. This lack of reproducibility increases the rate of complications from over-or undertreatment and inhibits determination of the optimal treatment endpoints for different retinal disorders. To this end, a feedback-controlled photocoagulator could make retinal photocoagulation a safer, more reproducible, and faster procedure. A dynamic confocal reflectometer was integrated into a slit lamp laser delivery system. Real-time reflectance changes on the retinas of pigmented rabbits were obtained by monitoring the increasing back-scattered light of the coagulating beam during argon laser photocoagulation. Reproducible temporal reflectance patterns were measured that correlated with ophthalmoscopically assessed lesion intensity independent of the exposure parameters, the transparency of the optical media, and the focusing conditions. As a step toward the development of a feedback-controlled photocoagulator, the confocal reflectometer has been proven in animal trials closely resembling clinical practice. © 1994 Wiley-Liss, Inc.

We investigated the use of picosecond Nd:YAG laser pulses for intraocular microsurgery. With a pulse duration of 100 picoseconds, only 70 μJ of pulse energy is required to consistently produce optical breakdown in the deep vitreous. This pulse energy is nearly two orders of magnitude less than the typical pulse energies used in conventional (nanosecond) photodisruptors. The reduced pulse energy results in a smaller zone of tissue damage, an important consideration when operating close to the retina or other sensitive ocular structures. Efficient cutting action is achieved by applying multiple pulses at a moderately high repetition rate of 50–200 Hz. An in vitro model was developed to assess axial confinement of picosecond photodisruption. In vivo vitreous membrane surgery was performed in experimental rabbit eyes to demonstrate a potential clinical application of picosecond laser—induced optical breakdown. © 1994 Wiley-Liss, inc.

Nd:YAG laser photodisruption with nanosecond (ns) pulses is an established method for intraocular surgery. In order to assess whether an increased precision can be achieved by the use of picosecond (ps) pulses, the plasma size, the shock wave characteristics, and the cavitation bubble expansion after optical breakdown with ps- and ns-laser pulses were investigated by time-resolved photography and acoustic measurements. Nd:YAG laser pulses with a duration of 30 ps and 6 ns, respectively, were focused into a water-filled glass cuvette. Frequency doubled light from the same laser pulses was optically delayed between 2 ns and 136 ns and used as illumination light source for photography. Since the individual events were well reproducible, the shock wave and bubble wall position could be determined as a function of time. From the slope of these r(t) curves, the shock wave and bubble wall velocities were determined, and the shock wave pressure was calculated from the shock velocity. The plasma size at various laser pulse energies was measured from photographs of the plasma radiation. The breakdown thresholds at 30 ps and 6 ns pulse duration were found to be 15 microJ and 200 microJ, respectively. At threshold, ps-plasmas are shorter than ns-plasmas, but at the same pulse energy they are always approximately 2.5 times longer. The initial shock pressures were 17 kbar after ps-pulses with an energy of 50 microJ, and 21 kbar after 1 mJ ns-pulses. The pressure amplitude decayed much faster after the ps-pulses. The maximum expansion velocity of the cavitation bubble was 350 m/s after a 50 microJ ps-pulse, but 1,600 m/s after a 1 mJ ns-pulse. The side effects of intraocular microsurgery associated with shock wave emission and cavitation bubble expansion can be considerably reduced by the use of ps-pulses, and new applications of photodisruption may become possible.

Retinal photocoagulation is an established treatment for various retinal diseases. The temperature development during a treatment can be monitored by applying short laser pulses in addition to the treatment laser light. The laser pulses induce optoacoustic pressure waves that can be detected at the cornea. Aim of this work is the investigation of the accuracy of the determined temperatures during a treatment. To calibrate the temperature dependency of the measured pressure, whole enucleated porcine eyes were heated using an infrared laser beam, while probing the retina optoacoustically. The temperatures and the optoacoustic pressure waves were measured simultaneously using thermocouples and a piezoelectric element, respectively. From the deviation of the individual measurements an error of less than 15% in the calibration regime between 37 °C to 55 °C was found. Furthermore, the spatial and temporal temperature course was investigated. Calculations were performed to simulate the temporal and spatial temperature development during photocoagulation. A theoretical model to determine the peak temperature of the irradiated tissue from the mean temperature measured by optoacoustics was developed. The validity of the model was experimentally examined by heating the retina of porcine eyes with a laser beam diameter of 500 µm while successively measuring the temperature optoacoustically with a probe beam diameter of 500 µm and 100 µm at the center of the heated area, respectively. The deviation of the theoretical model and the experimental results were found to be less than 7µ.

Selective retina treatment (SRT) is a laser based method to treat retinal diseases associated with disorders of the retinal pigment epithelium (RPE) while preserving photoreceptors and choroid. Applying microsecond laser pulses to the 100- 200 strongly absorbing melanin granules inside the RPE cells induces transient micro bubbles which disrupt the cells. Aim of this work is to understand bubble dynamics in clusters with respect to the influence of the adjacent retina. Bubble dynamics were investigated in vitro on porcine RPE. An about 200 μm thick layer of agarose gel was applied to the RPE layer in order to simulate the mechanical properties of retina. Different laser pulse durations from 1 ns (532 nm, Nd:YAG) to 1.7 μs (527 nm, Nd:YLF) were used. The bubbles were investigated interferometrically (fiber interferometer @ 830 nm) and with fast flash photography (25 ns flash duration). Bubble lifetimes were measured. The results show that with retina phantoms the bubble formation threshold was reached at 2.5 times higher irradiation than without retina phantom for 1.7 μs laser pulses. The microbubbles generated with 1 ns laser pulses were almost not influenced by the agarose layer. Irradiation twofold over bubble formation threshold resulted in 3.5 times longer bubble lifetimes for μs and 2 times longer for ns pulse durations, respectively.

Retinal photocoagulation is a long time established treatment for a variety of retinal diseases, most commonly applied for diabetic macular edema and diabetic retinopathy. The damage extent of the induced thermal coagulations depend on the temperature increase and the time of irradiation. So far, the induced temperature rise is unknown due to intraocular variations in light transmission and scattering and RPE/choroidal pigmentation, which can vary inter- and intraindividually by more than a factor of four. Thus in clinical practice, often stronger and deeper coagulations are applied than therapeutically needed, which lead to extended retinal damage and strong pain perception. The final goal of this project focuses on a dosimetry control, which automatically generates a desired temperature profile and thus coagulation strength for every individual coagulation spot, ideally unburden the ophthalmologist from any laser settings. In this paper we present the first realtime temperature measurements achieved on patients during retinal photocoagulation by means of an optoacoustic method, making use of the temperature dependence of the thermal expansion coefficient of retinal tissue. Therefore, nanosecond probe laser pulses are repetitively and simultaneously applied with the treatment radiation in order to excite acoustic waves, which are detected at the cornea with an ultrasonic transducer embedded in the contact lens and then are processed by PC.

Retinal laser photocoagulation is an established treatment method for many retinal diseases like macula edema or diabetic retinopathy. The selection of the laser parameters is so far based on post treatment evaluation of the lesion size and strength. Due to local pigment variations in the fundus and individual transmission the same laser parameters often lead to an overtreatment. Optoacoustic allows a non invasive monitoring of the retinal temperature increase during retinal laser irradiation by measuring the temperature dependent pressure amplitudes, which are induced by short probe laser pulses. A 75 ns/ 523 nm Nd:YLF was used as a probe laser at a repetition rate of 1 kHz, and a cw / 532 nm treatment laser for heating. A contact lens was modified with a ring-shaped ultrasonic transducer to detect the pressure waves at the cornea. Temperatures were collected for irradiations leading to soft or invisible lesions. Based on this data the threshold for denaturation was found. By analyzing the initial temperature increase, the further temperature development during irradiation could be predicted. An algorithm was found to calculate the irradiation time, which is needed for a soft lesion formation, from the temperature curve. By this it was possible to provide a real-time dosimetry by automatically switching off the treatment laser after the calculated irradiation time. Automatically controlled coagulations appear softer and more uniformly.

Repetitive exposure to short laser pulses is shown to cause selective damage to absorbing structures (cells, organelles, or enzymes) with pulse energies below the threshold energy for single-pulse damage. Directly adjacent structures are spared in vivo. Additivity of (presumably nonphotochemical) subthreshold effects is demonstrated. Selective damage to the retinal pigment epithelium with sparing of the neural retina is shown (514 nm, 5 microseconds, 1-500 pulses at 500 Hz, 2- to 10-microJ pulse energy). A melanin granule model has been developed and applied to the experimental situation. Histological results as well as the basic mechanism for these effects are discussed.