Boris Považay

Ultrahigh resolution optical coherence tomography and pancorrection for cellular imaging of the living human retina

Cellular in vivo visualization of the three dimensional architecture of individual human foveal cone photoreceptors is demonstrated by combining ultrahigh resolution optical coherence tomography and a novel adaptive optics modality. Isotropic resolution in the order of 2-3 microm, estimated from comparison with histology, is accomplished by employing an ultrabroad bandwidth Titanium:sapphire laser with 140 nm bandwidth and previous correction of chromatic and monochromatic ocular aberrations. The latter, referred to as pancorrection, is enabled by the simultaneous use of a specially designed lens and an electromagnetically driven deformable mirror with unprecedented stroke for correcting chromatic and monochromatic aberrations, respectively. The increase in imaging resolution allows for resolving structural details of distal elements of individual foveal cones: inner segment zones--myoids and ellipsoids--are differentiated from outer segments protruding into pigment epithelial processes in the retina. The presented technique has the potential to unveil photoreceptor development and pathogenesis as well as improved therapy monitoring of numerous retinal diseases.

Mapping Choroidal and Retinal Thickness Variation in Type 2 Diabetes using Three-Dimensional 1060-nm Optical Coherence Tomography

PURPOSE To map choroidal (ChT) and retinal thickness (RT) in healthy subjects and patients with diabetes with and without maculopathy using three dimensional 1060-nm optical coherence tomography (3D-1060nm-OCT). METHODS Sixty-three eyes from 42 diabetic subjects (41-82 years of age; 11 females) grouped according to a custom scheme using Early Treatment Diabetic Retinopathy Study definitions for pathology within 1 disc-diameter of fovea (without pathology [NDR], microaneurysms [M1], exudates [M2], clinically significant macular edema [CSME]) and 16 eyes from 16 healthy age matched subjects (38-79 years of age; 11 females) were imaged by 3D-1060nm-OCT performed over a 36° × 36° field of view. Axial length, 45° fundus photographs, body mass index, plasma glucose, and blood pressure measurements were recorded. The ChT at the subfoveal location and ChT maps between RPE and the choroidal-scleral interface were generated and statistically analyzed. RESULTS RT maps show thinning in the NDR group but an increase in thickness with increasing maculopathy in the temporal and central regions (unpaired t-test; P < 0.05). ChT mapping of all diabetic patients revealed central and inferior thinning compared to healthy eyes (unpaired t-test; P < 0.001). Subfoveal ChT (mean ± SD) for healthy eyes was 327 ± 74 μm, which was significantly thicker than all diabetic groups (214 ± 55 μm for NDR, 208 ± 49 μm for M1, 205 ± 54 μm for M2, and 211 ± 76 μm for CSME (ANOVA P < 0.001; Tukey P < 0.001). CONCLUSIONS 3D-1060nm-OCT has shown that the central choroid is thinner in all type 2 diabetic eyes regardless of disease stage. The choroidal thinning may exceed the magnitude of possible choriocapillaris atrophy. In contrast to the conventional assessment of pathologic thickness change in several locations, thickness maps allow investigation of the choroid over the extent of affected areas.

Adaptive optics optical coherence tomography at 120,000 depth scans/s for non-invasive cellular phenotyping of the living human retina

This paper presents a successful combination of ultra-high speed (120,000 depth scans/s), ultra-high resolution optical coherence tomography with adaptive optics and an achromatizing lens for compensation of monochromatic and longitudinal chromatic ocular aberrations, respectively, allowing for non-invasive volumetric imaging in normal and pathologic human retinas at cellular resolution. The capability of this imaging system is demonstrated here through preliminary studies by probing cellular intraretinal structures that have not been accessible so far with in vivo, non-invasive, label-free imaging techniques, including pigment epithelial cells, micro-vasculature of the choriocapillaris, single nerve fibre bundles and collagenous plates of the lamina cribrosa in the optic nerve head. In addition, the volumetric extent of cone loss in two colour-blinds could be quantified for the first time. This novel technique provides opportunities to enhance the understanding of retinal pathogenesis and early diagnosis of retinal diseases.

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.

Dispersion encoded full range frequency domain optical coherence tomography

We propose an iterative algorithm that exploits the dispersion mismatch between reference and sample arm in frequency-domain optical coherence tomography (FD-OCT) to effectively cancel complex conjugate mirror terms in individual A-scans and thereby generate full range tomograms. The resulting scheme, termed dispersion encoded full range (DEFR) OCT, allows distinguishing real structures from complex conjugate mirror artifacts. Even though DEFR-OCT has higher post-processing complexity than conventional FD-OCT, acquisition speed is not compromised since no additional A-scans need to be measured, thereby rendering this technique robust against phase fluctuations. The algorithm uses numerical dispersion compensation and exhibits similar resolution as standard processing. The residual leakage of mirror terms is further reduced by incorporating additional knowledge such as the power spectrum of the light source. The suppression ratio of mirror signals is more than 50 dB and thus comparable to complex FD-OCT techniques which use multiple A-scans.

Full-field time-encoded frequency-domain optical coherence tomography

Ultrahigh axial resolution surface profiling as well as volumetric optical imaging based on time encoded optical coherence tomography in the frequency domain without any mechanical scanning element is presented. A frequency tuned broad bandwidth titanium sapphire laser is interfaced to an optical microscope (Axioskop 2 MAT, Carl Zeiss Meditec) that is enhanced with an interferometric imaging head. The system is equipped with a 640 x 480 pixel CMOS camera, optimized for the 800 nm wavelength tuning range for transmission and reflection measurements of a microscopic sample. Sample volume information over 1.3 x 1 x 0.2 mm(3) with ~3 mum axial and ~4 mum transverse resolution in tissue is acquired by a single wavelength scan over more than 100 nm optical bandwidth from <760 to >860 nm with 128-2048 equidistant optical frequency steps with an acquisition time of 1 to 50 ms per step. Topography and tomography with a signal to noise ratio of 83 dB is demonstrated on test surfaces and biological specimen respectively. This novel OCT technique promises to enable high speed, three dimensional imaging by employing high frame rate cameras and state of the art tunable lasers in a mechanically stable environment, due to lack of moving components while reducing the intensity on the sample.

Chromatic aberration correction of the human eye for retinal imaging in the near infrared

An achromatizing lens has been designed for the human eye in the near infrared range, from 700 to 900 nm, for retinal imaging purposes. Analysis of the performance of the lens, including tolerance to misalignments, has been mathematically accomplished by using an existing eye model. The calculations have shown a virtually perfect correction of the ocular longitudinal chromatic aberration, while still keeping a high optical quality. Ocular aberrations in five subjects have been measured with and without the achromatizing lens by using a Hartmann-Shack wavefront sensor and a broad bandwidth femtosecond Ti:sapphire laser in the spectral range of interest with a set of interference filters, studying the benefits and limits in the use of the achromatizing lens. Ocular longitudinal chromatic aberration has been experimentally demonstrated to be fully corrected by the proposed lens, with no induction of any other parasitic aberration. The practical implementation of the achromatizing lens for Ophthalmoscopy, specifically for optical coherence tomography where the use of polychromatic light sources in the near infrared portion of the spectrum is mandatory, has been considered. The potential benefits of using this lens in combination with adaptive optics to achieve a full aberration correction of the human eye for retinal imaging have also been discussed.

Choroidal thinning in diabetes type 1 detected by 3-dimensional 1060 nm optical coherence tomography.

PURPOSE To map choroidal (ChT) and retinal thickness (RT) in patients with diabetes type 1 with and without maculopathy and retinopathy in order to compare them with healthy subjects using high speed 3-dimensional (3D) 1060 nm optical coherence tomography (OCT). METHODS Thirty-three eyes from 33 diabetes type 1 subjects (23-57 years, 15 male) divided into groups of without pathology (NDR) and with pathology (DR; including microaneurysms, exudates, clinically significant macular-oedema and proliferative retinopathy) were compared with 20 healthy axial eye length and age-matched subjects (24-57 years, 9 male), imaged by high speed (60.000 A-scans/s) 3D 1060 nm OCT performed over 36° × 36° field of view. Ocular health status, disease duration, body mass index, haemoglobin-A1c, and blood pressure (bp) measurements were recorded. Subfoveal ChT, and 2D topographic maps between retinal pigment epithelium and the choroidal/scleral-interface, were automatically generated and statistically analyzed. RESULTS Subfoveal ChT (mean ± SD, μm) for healthy eyes was 388 ± 109; significantly thicker than all diabetic groups, 291 ± 64 for NDR, and 303 ± 82 for DR (ANOVA P < 0.004, Tukey P = 0.01 for NDR and DR). Thinning did not relate to recorded factors (multi-regression analysis, P > 0.05). Compared with healthy eyes and the NDR, the averaged DR ChT-map demonstrated temporal thinning that extended superiorly and temporal-inferiorly (unpaired t-test, P < 0.05). Foveal RT and RT-maps showed no statistically significant difference between groups (mean SD, μm, healthy 212 ± 17, NDR 217 ± 15, DR 216 ± 27, ANOVA P > 0.05). CONCLUSIONS ChT is decreased in diabetes type 1, independent of the absence of pathology and of diabetic disease duration. In eyes with pathology, 3D 1060 nm OCT averaged maps showed an extension of the thinning area matching retinal lesions and suggesting its involvement on onset or progression of disease.

In vivo ultrahigh-resolution optical coherence tomography of mouse colon with an achromatized endoscope.

Endoscopic ultrahigh-resolution optical coherence tomography (OCT) enables collection of minimally invasive cross-sectional images in vivo, which may be used to facilitate rapid development of reliable mouse models of colon disease as well as assess chemopreventive and therapeutic agents. The small physical scale of mouse colon makes light penetration less problematic than in other tissues and high resolution acutely necessary. In our 2-mm diameter endoscopic time domain OCT system, isotropic ultrahigh-resolution is supported by a center wavelength of 800 nm and full-width-at-half-maximum bandwidth of 150 nm (mode-locked titanium:sapphire laser) combined with 1:1 conjugate imaging of a small core fiber. A pair of KZFSN5/SFPL53 doublets provides excellent color correction to support wide bandwidth throughout the imaging depth. A slight deviation from normal beam exit angle suppresses collection of the strong back reflection at the exit window surface. Our system achieves axial resolution of 3.2 microm in air and 4.4-microm lateral spot diameter with 101-dB sensitivity. Microscopic features too small to see in mouse tissue with conventional resolution systems, including colonic crypts, are clearly resolved. Resolution near the cellular level is potentially capable of identifying abnormal crypt formation and dysplastic cellular organization.

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 (Si3N4) and silicon dioxide (SiO2) 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, Si3N4/SiO2 waveguides have a potential to serve as a platform for passive photonic integrated circuits for OCT.