Photonic integrated Mach-Zehnder interferometer with an on-chip reference arm for optical coherence tomography

Optical coherence tomography (OCT) is a noninvasive, three-dimensional imaging modality with several medical and industrial applications. Integrated photonics has the potential to enable mass production of OCT devices to significantly reduce size and cost, which can increase its use in established fields as well as enable new applications. Using silicon nitride (Si(3)N(4)) and silicon dioxide (SiO(2)) waveguides, we fabricated an integrated interferometer for spectrometer-based OCT. The integrated photonic circuit consists of four splitters and a 190 mm long reference arm with a foot-print of only 10 × 33 mm(2). It is used as the core of a spectral domain OCT system consisting of a superluminescent diode centered at 1320 nm with 100 nm bandwidth, a spectrometer with 1024 channels, and an x-y scanner. The sensitivity of the system was measured at 0.25 mm depth to be 65 dB with 0.1 mW on the sample. Using the system, we imaged human skin in vivo. With further optimization in design and fabrication technology, Si(3)N(4)/SiO(2) waveguides have a potential to serve as a platform for passive photonic integrated circuits for OCT

Real-time OCT feedback-controlled RPE photodisruption in ex vivo porcine eyes using 8 microsecond laser pulses

Selective retinal pigment epithelium (RPE) photodisruption requires reliable real-time feedback dosimetry (RFD) to prevent unwanted overexposure. In this study, optical coherence tomography (OCT) based RFD was investigated in ex vivo porcine eyes exposed to laser pulses of 8 µs duration (wavelength: 532 nm, exposure area: 90 × 90 µm2, radiant exposure: 247 to 1975 mJ/µm2). For RFD, fringe washouts in time-resolved OCT M-scans (central wavelength: 870 nm, scan rate: 85 kHz) were compared to an RPE cell viability assay. Statistical analysis revealed a moderate correlation between RPE lesion size and applied treatment energy, suggesting RFD adaptation to inter- and intraindividual RPE pigmentation and ocular transmission

Retinal and choroidal thickness in early age-related macular degeneration

Purpose: To compare retinal thickness and choroidal thickness at increasing retinal eccentricity in individuals with early age-related macular degeneration (AMD) and in healthy controls using enhanced choroidal penetration, 3-dimensional optical coherence tomography at 1060 nm. Design: Cross-sectional study. Methods: Individuals with early AMD (n = 16; mean age, 71.6 ± 8.5 years) and a comparison group of healthy controls (n = 16; 67.6 ± 5.4 years) were recruited. Three-dimensional (20 degrees × 20 degrees) long-wavelength optical coherence tomography (1060 nm) images (approximately 8-μm axial resolution; 47 000 A scans/second, centered on the fovea) were obtained from all participants after pupil dilation. Retinal thickness was measured between the inner limiting membrane and the retinal pigment epithelium. Choroidal thickness was measured between the retinal pigment epithelium and the choroid–scleral interface. Thickness measurements were obtained subfoveally and at 0.5-mm intervals to a maximum of 2.0 mm nasally, temporally, superiorly, and inferiorly. The main outcome measures were retinal and choroidal thickness (measured in micrometers) at different eccentricities on vertical and horizontal meridians. Results: Mean retinal thickness was reduced significantly in the group of participants with early AMD compared with the control group at multiple locations within 2.0 mm of the fovea. This difference was most significant at the fovea, where the mean retinal thickness of the early AMD group was 179 ± 27 μm and that of the control group was 202 ± 18 μm (P = .008). There was no significant difference in choroidal thickness between groups at any location. Conclusions: Retinal thickness is reduced in early AMD, but choroidal thickness seems to be unaffected by the early disease process

Dynamic OCT Signal Loss for Determining RPE Radiant Exposure Damage Thresholds in Microsecond Laser Microsurgery

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

Real-Time Optical Coherence Tomography Controlled Microsecond Laser Retinal Microsurgery: First In-vivo Results

Reliable mild photocoagulation and selective retina therapy (SRT) selectively damaging the retinal pigment epithelium (RPE) while sparing the neuroretina, the photoreceptors as well as the choroid are highly demanded. However, due to the inter- and intraindividual variability of RPE and choroidal absorption, optical microsurgery requires reliable real-time laser dosing to prevent unwanted overexposure and extended damage of the neuroretina. In this experiment optical coherence tomography (OCT) was implemented to detect minimal damage, and a laser feedback control algorithm was used for real-time dosing. For the first time in-vivo experiments on rabbits were performed with microsecond laser pulses of varying duration. Pigment rabbit eyes (n=6) were exposed to laser pulses of 4, 8, 12, and 20 μs in duration (wavelength, 532 nm; ramp-mode, maximum 15 pulses; repetition rate, 100 Hz). Therefore, a system with a scanning laser ophthalmoscope and spectral-domain OCT (Heidelberg Engineering) extended with a prototype laser (Meridian Medical) was used. For each laser lesion, the increasing ramp’s end energy was individually controlled in real-time using OCT dosimetry (central wavelength, 870 nm; scan rate, 80 kHz). Within 1 hour after irradiation, retinal changes were assessed with fluorescein angiography (FA), indocyanine green angiography (ICGA), color fundus photography (CFP) and OCT. OCT dosimetry utilizing the control algorithm can interrupt the ramp-mode in real-time for each lesion individually. The preconditioned algorithm enabled treatment with a clearly visible breakdown of the blood-retinal barrier (BRB) according to FA and ICGA imaging and barely visible treatment lesions according to CFP. OCT B-scans through the treated areas provided a first indication of the morphological tissue impact. Preliminary evaluation shows that the algorithm stopped the laser at 4 μs at a ramp end energy of 53 μJ (corresponds to 13/15 pulses), at 8 μs at 68 μJ (5/15 pulses), at 12 μs at 74 μJ (7/15 pulses), and at 20 μs at 100 μJ (1/15 pulses). The novel system with OCT based laser dosing proved to induce minimal visible damage and BRB breakdown in a wide range of pulse durations. The new irradiation scheme and algorithm are being optimized and tested in multiple subjects to further limit unwanted damage and enable pure RPE selective laser microsurgery in real-time. This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually

Short-term follow up after Large-Area RPE Removal by Microsecond Laser followed by hiPS-RPE suspension transplantation in rabbits

Cell therapy is a promising treatment for retinal pigment epithelium (RPE)-associated eye diseases. Herein, microsecond laser irradiation targeting RPE cells was used for large-area RPE removal followed by subretinal injection of human induced pluripotent stem cell derived RPE (hiPS-RPE). 19 immunosuppressed pigmented rabbits (Chinchilla bastard hybrid) underwent a large area RPE removal using an infrared reflectance (IR) confocal scanning laser ophthalmoscope (cSLO) with spectral-domain optical coherence tomography (SD-OCT) system (Heidelberg Engineering ) extended with a prototype laser (modified Merilas 532 shortpulse ophthalmic laser photocoagulator, Meridian Medical) (wavelength, 532 nm; pulse duration, 8 µs), followed by a 25G vitrectomy. Subsequently, a suspension of hiPS-RPE (1000 cells/ µl) was grafted subretinally into the RPE laser lesion under real-time intraoperative OCT imaging (RESCAN 700, Zeiss) by manual injection via a 25/38G cannula connected to a 100µl Hamilton syringe. 5 rabbits served as a control with hiPS-RPE injected subretinally over healthy RPE. The rabbits were followed with in vivo multimodal retinal imaging at baseline after laser and then for 7 days including fluorescein (FA) and indocyanine angiography (ICGA), aw well as SD-OCT (Spectralis ®, Heidelberg Engineering). Baseline imaging of RPE laser wounds showed mild late phase FA/ICGA leakage, with normal outer retinal and choroidal reflectivity on OCT, without signs of coagulation. The size of the RPE wounds was typically 10-12mm2. Real time iOCT showed a directed spread of the bleb retinal detachment (bRD) within the lasered zone, in contrast to a circular spread in controls. Subretinal injection ranged from 5-20µl, with lesser volumes/ larger bRD areas over lasered regions. At 7 days, implanted regions showed FA/ICGA leakage, blockage due to hyperpigmentation was observed mostly at the edges of the lasered zone; OCT showed hyperreflectivity of the outer retina with RPE irregularities. Control implantation sites showed hyperreflectivity in all retinal layers and a variably thickened RPE band suggesting clumping. Microsecond laser irradiation to the RPE seems to accelerate the subretinal integration of hiPS-RPE, when compared to subretinal injection over intact RPE. Future work will address correlation of multimodal imaging and histology. This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually

Selective Large-Area Retinal Pigment Epithelial Removal by Microsecond Laser in Preparation for Cell Therapy

Purpose: Cell therapy is a promising treatment for retinal pigment epithelium (RPE)- associated eye diseases such as age-related macular degeneration. Herein, selective microsecond laser irradiation targeting RPE cells was used for minimally invasive, large- area RPE removal in preparation for delivery of retinal cell therapeutics. Methods: Ten rabbit eyes were exposed to laser pulses 8, 12, 16, and 20 μs in duration (wavelength, 532 nm; top-hat beam profile, 223 × 223 μm2). Post-irradiation retinal changes were assessed with fluorescein angiography (FA), indocyanine green angiogra- phy (ICGA), and optical coherence tomography (OCT). RPE viability was evaluated with an angiographic probit model. Following vitrectomy, a subretinal injection of balanced salt solution was performed over a lasered (maximum 13.6 mm2) and untreated control area. Bleb retinal detachment (bRD) morphology was then evaluated by intraoperative OCT. Results: Within 1 hour after irradiation, laser lesions showed FA and ICGA leakage. OCT revealed that large-area laser damagewas limited to the RPE. The angiographic median effective dose irradiation thresholds (ED50)were45μJ(90 mJ/cm2)at8μs,52μJ(104 mJ/cm2) at 12 μs, 59 μJ (118 mJ/cm2)at16μs,and71μJ(142mJ/cm2) at 20 μs. Subretinal injection over the lasered area resulted in a controlled, shallowbRD rise, whereas control blebs were convex in shape, with less predictable spread. Conclusions: Large-area, laser-based removal ofhost RPEwithout visible photoreceptor damage is possible and facilitates surgical retinal detachment. Translational Relevance: Selective microsecond laser-based, large-area RPE removal prior to retinal cell therapy may reduce iatrogenic trauma. Introductio

Optical coherence tomography controlled selective retina therapy with a novel microsecond laser

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