Depixelation of coherent fiber bundle imaging by fiber-core-targeted scanning.

A novel fast proximal scanning method, to the best of our knowledge, termed fiber-core-targeted scanning (FCTS), is proposed for illuminating individual fiber cores sequentially to remove the pixelation effect in fiber bundle (FB) imaging. FCTS is based on a galvanometer scanning system. Through a dynamic control of the scan trajectory and speed using the prior knowledge of fiber core positions, FCTS experimentally verifies a precise sequential delivery of laser pulses into fiber cores at a maximal speed of 45,000 cores per second. By applying FCTS on a FB-based photoacoustic forward-imaging probe, the results demonstrate that FCTS eliminates the pixelation effect and improves the imaging quality.

Functional optical coherence tomography and photoacoustic microscopy imaging for zebrafish larvae.

We present a dual modality functional optical coherence tomography and photoacoustic microscopy (OCT-PAM) system. The photoacoustic modality employs an akinetic optical sensor with a large imaging window. This imaging window enables direct reflection mode operation, and a seamless integration of optical coherence tomography (OCT) as a second imaging modality. Functional extensions to the OCT-PAM system include Doppler OCT (DOCT) and spectroscopic PAM (sPAM). This functional and non-invasive imaging system is applied to image zebrafish larvae, demonstrating its capability to extract both morphological and hemodynamic parameters in vivo in small animals, which are essential and critical in preclinical imaging for physiological, pathophysiological and drug response studies.

Comprehensive vascular imaging using optical coherence tomography-based angiography and photoacoustic tomography.

Studies have proven the relationship between cutaneous vasculature abnormalities and dermatological disorders, but to image vasculature noninvasively

Phase-stable swept source OCT angiography in human skin using an akinetic source.

We demonstrate noninvasive structural and microvascular contrast imaging of human skin in vivo, using phase difference swept source OCT angiography (pOCTA). The pOCTA system employs an akinetic, all-semiconductor, highly phase-stable swept laser source which operates at 1340 nm central wavelength, with 37 nm bandwidth (at 0 dB region) and 200 kHz A-scan rate. The phase sensitive detection does not need any external phase stabilizing implementations, due to the outstanding high phase linearity and sweep phase repeatability within 2 mrad. We compare the performance of phase based OCTA to speckle based OCTA for visualizing human vascular networks. pOCTA shows better contrast especially for deeper vascular details as compared to speckle based OCTA. The phase stability of the akinetic source allows the OCTA system to show decent vascular contrast only with 2 B-scans. We compare the performance of using 2 versus 4 B-scans for calculating the vascular contrast. Finally, the performance of a 100 nm bandwidth akinetic laser at 1310 nm is investigated for both OCT and OCTA.

High-speed polarization sensitive optical coherence tomography scan engine based on Fourier domain mode locked laser.

We report on a new swept source polarization sensitive optical coherence tomography scan engine that is based on polarization maintaining (PM) fiber technology. The light source is a Fourier domain mode locked laser with a PM cavity that operates in the 1300 nm wavelength regime. It is equipped with a PM buffer stage that doubles the fundamental sweep frequency of 54.5 kHz. The fiberization allows coupling of the scan engine to different delivery probes. In a first demonstration, we use the system for imaging human skin at an A-scan rate of 109 kHz. The system illuminates the sample with circularly polarized light and measures reflectivity, retardation, optic axis orientation, and Stokes vectors simultaneously. Furthermore, depolarization can be quantified by calculating the degree of polarization uniformity (DOPU). The high scanning speed of the system enables dense sampling in both, the x- and y-direction, which provides the opportunity to use 3D evaluation windows for DOPU calculation. This improves the spatial resolution of DOPU images considerably.

Polarization sensitive optical coherence tomography of melanin provides intrinsic contrast based on depolarization.

Polarization sensitive optical coherence tomography (PS-OCT) is a functional extension of OCT. In addition to imaging based on tissue reflectivity, PS-OCT also enables depth-resolved mapping of sample polarization properties such as phase-retardation, birefringent axis orientation, Stokes vectors, and degree of polarization uniformity (DOPU). In this study, PS-OCT was used to investigate the polarization properties of melanin. In-vitro measurements in samples with varying melanin concentrations revealed polarization scrambling, i.e. depolarization of backscattered light. Polarization scrambling in the PS-OCT images was more pronounced for higher melanin concentrations and correlated with the concentration of the melanin granules in the phantoms. Moreover, in-vivo PS-OCT was performed in the retinas of normal subjects and individuals with albinism. Unlike in the normal eye, polarization scrambling in the retinal pigment epithelium (RPE) was less pronounced or even not observable in PS-OCT images of albinos. These results indicate that the depolarizing appearance of pigmented structures like, for instance, the RPE is likely to be caused by the melanin granules contained in these cells.

Speckle noise reduction in high speed polarization sensitive spectral domain optical coherence tomography.

We present a high speed polarization sensitive spectral domain optical coherence tomography system based on polarization maintaining fibers and two high speed CMOS line scan cameras capable of retinal imaging with up to 128 k A-lines/s. This high imaging speed strongly reduces motion artifacts and therefore averaging of several B-scans is possible, which strongly reduces speckle noise and improves image quality. We present several methods for averaging retardation and optic axis orientation, the best one providing a 5 fold noise reduction. Furthermore, a novel scheme of calculating images of degree of polarization uniformity is presented. We quantitatively compare the noise reduction depending on the number of averaged frames and discuss the limits of frame numbers that can usefully be averaged.

In vivo investigation of human cone photoreceptors with SLO/OCT in combination with 3D motion correction on a cellular level.

We present a further improvement of our SLO/OCT imaging system which enables to practically eliminate all eye motion artifacts with a correction accuracy approaching sub-cellular dimensions. Axial eye tracking is achieved by using a hardware based, high speed tracking system that consists of a rapid scanning optical delay line in the reference arm of the interferometer. A software based algorithm is employed to correct for transverse eye motion in a post-processing step. The instrument operates at a frame rate of 40 en-face fps with a field of view of approximately 1 degrees x 1 degrees. Dynamic focusing enables the recording of 3D volumes of the human retina with cellular resolution throughout the entire imaging depth. Several volumes are stitched together to increase the total field of view. Different features of the three dimensional structure of cone photoreceptors are investigated in detail and at different eccentricities from the fovea.

Segmentation and quantification of retinal lesions in age-related macular degeneration using polarization-sensitive optical coherence tomography.

We present polarization-sensitive optical coherence tomography (PS-OCT) for quantitative assessment of retinal pathologies in age-related macular degeneration (AMD). On the basis of the polarization scrambling characteristics of the retinal pigment epithelium, novel segmentation algorithms were developed that allow one to segment pathologic features such as drusen and atrophic zones in dry AMD as well as to determine their dimensions. Results from measurements in the eyes of AMD patients prove the ability of PS-OCT for quantitative imaging based on the retinal features polarizing properties. Repeatability measurements were performed in retinas diagnosed with drusen and geographic atrophy in order to evaluate the performance of the described methods. PS-OCT appears as a promising imaging modality for three-dimensional retinal imaging and ranging with additional contrast based on the structures' tissue-inherent polarization properties.

Phase contrast coherence microscopy based on transverse scanning.

We present what we believe to be a novel approach to measuring optical path length differences with a precision of a few nanometers. The instrument is based on transverse scanning or en-face optical coherence tomography. Owing to the fast motion of the scanning beam over the sample, excellent phase stability in the transverse direction is achieved. Hence, phase changes caused by the varying optical path lengths within the sample arm occur with high frequency in the fast scanning direction. These changes are well separated from the rather slow phase changes introduced by jitter within the interferometer and can therefore be measured. The en-face imaging speed of the instrument is 40 fps (520 x 200 pixels). The measured precision of the method to detect small changes in optical path lengths was approximately 3 nm.

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.

Serial endoscopy in azoxymethane treated mice using ultra-high resolution optical coherence tomography.

PURPOSE: Optical coherence tomography (OCT) is a minimally invasive, depth-resolved imaging tool that can be implemented in a small diameter endoscope for imaging mouse models of colorectal cancer (CRC). In this study, we utilized ultrahigh resolution (UHR) OCT to serially image the lower colon of azoxymethane (AOM) treated A/J mouse models of CRC in order to monitor the progression of neoplastic transformations and determine if OCT is capable of identifying early disease. EXPERIMENTAL DESIGN: Thirteen AOM treated A/J and two control A/J mice were surveyed at four timepoints (8, 14, 22 and 26 weeks post AOM treatment) using a 2.0 mm diameter UHR OCT endoscopic system with 3.2 microm axial and 4.4 microm lateral resolution. Histological samples obtained at the final timepoint served as the diagnostic reference. A blinded expert panel of mouse colon pathologists provided diagnoses from the OCT images based on criteria developed from a separate training set of OCT images. Panel results were compared to histological diagnoses assigned by a blinded pathologist. RESULTS: At the final imaging timepoint, 95% of adenomas and 23% of gastrointestinal neoplasias (38% protruding GINs and 9% non-protruding GINs) were correctly diagnosed. The panel identified 68% of disease foci (95% adenoma, 76% protruding GINs and 13% non-protruding GINs). Over the OCT imaging timepoints, disease progression followed a typical succession, with normal or GIN preceding adenoma. CONCLUSIONS: Endoscopic UHR OCT enabled accurate diagnosis of adenomas, identification of protruding GIN and non-destructive visualization of CRC progression, providing a tool for cancer research in animal models.

Three-dimensional optical coherence tomography at 1050 nm versus 800 nm in retinal pathologies: enhanced performance and choroidal penetration in cataract patients.

Frequency domain optical coherence tomography (FD-OCT), based on an all-reflective high-speed InGaAs spectrometer, operating in the 1050 nm wavelength region for retinal diagnostics, enables high-speed, volumetric imaging of retinal pathologies with greater penetration into choroidal tissue is compared to conventional 800 nm three-dimensional (3-D) ophthalmic FD-OCT systems. Furthermore, the lower scattering at this wavelength significantly improves imaging performance in cataract patients, thereby widening the clinical applicability of ophthalmic OCT. The clinical performance of two spectrometer-based ophthalmic 3-D OCT systems compared in respect to their clinical performance, one operating at 800 nm with 150 nm bandwidth (approximately 3 microm effective axial resolution) and the other at 1050 nm with 70 nm bandwidth (approximately 7 microm effective axial resolution). Results achieved with 3-D OCT at 1050 nm reveal, for the first time, decisive improvements in image quality for patients with retinal pathologies and clinically significant cataract.

Simultaneous SLO/OCT imaging of the human retina with axial eye motion correction.

It has been shown that transversal scanning (or en-face) optical coherence tomography (TS-OCT) represents an imaging modality capable to record high isotropic resolution images of the human retina in vivo. However, axial eye motion still remains a challenging problem of this technique. In this paper we introduce a novel method to compensate for this eye motion. An auxiliary spectral domain partial coherence interferometer (SD-PCI) was integrated into an existing TS-OCT system and used to measure accurately the position of the cornea. A light source emitting at 1310nm was used in the additional interferometer which enabled a nearly loss free coupling of the two measurement beams via a dichroic mirror. The recorded corneal position was used to drive an additional voice coil translation stage in the reference arm of the TS-OCT system to correct for axial eye motion. Currently, the correction can be performed with an update rate of ~200Hz. The TS-OCT instrument is operated with a line scan rate of 4000 transversal lines per second which enables simultaneous SLO/OCT imaging at a frame rate of 40fps. 3D data of the human retina with isotropic high resolution, that was sufficient to visualize the human cone mosaic in vivo, is presented.

Ultrahigh resolution optical coherence tomography of macular holes.

PURPOSE: To evaluate ultrahigh resolution optical coherence tomography (UHR OCT) for visualization of intraretinal layers, especially the photoreceptor inner segment and outer segment layers, in eyes with macular holes and after surgical intervention. METHODS: An UHR OCT system based on a titanium:sapphire laser was used, enabling in vivo cross-sectional retinal imaging with 3-micro m axial resolution. Typical, representative tomograms of 5 of 48 eyes from 36 patients demonstrated the potential of UHR OCT to detect morphologic changes in different stages of full-thickness macular holes and changes induced by surgical intervention. RESULTS: UHR OCT could detect subtle intraretinal changes in macular hole formation. Unprecedented visualization of photoreceptor impairment was achieved that appeared to be more extensive than the hole itself. Postoperatively, clinically closed holes showed restoration of the photoreceptor inner and outer segment layers of various extents, with residual disease in some eyes. CONCLUSION: In macular holes, UHR OCT allows for detection of even small morphologic changes of the retinal layers, especially the photoreceptor inner and outer segment layers. Therefore, it also represents a superior method to monitor the effect of surgical interventions. Preoperative photoreceptor impairment and the degree of postoperative restoration could possibly be associated with visual function. Hence, UHR OCT could lead to better understanding of macular hole pathogenesis and to more accurate disease prognosis.

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.

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.

Ultrahigh resolution optical coherence tomography in macular dystrophy.

PURPOSE: To visualize and investigate intraretinal changes in macular dystrophies with ultrahigh resolution optical coherence tomography (UHR OCT). DESIGN: Prospective observational case series. METHODS: setting: Department of Ophthalmology and Center for Biomedical Engineering and Physics, Christian Doppler Laboratory, Medical University of Vienna, Vienna, Austria. patients: Thirteen patients (23 eyes) with adult-onset foveomacular vitelliform dystrophy (AOFVD) and 14 patients (27 eyes) with Stargardt's disease (SD) or fundus flavimaculatus (FF). OBSERVATIONS: Imaging using a compact, new generation UHR OCT system, achieving considerably improved visualization of intraretinal layers, especially the photoreceptor layer. main outcome measures: UHR OCT tomograms visualizing intraretinal differences in morphology of AOFVD and SD/FF as location and extension of deposits and loss of photoreceptors. Central foveal thickness defined as distance between internal limiting membrane and photoreceptors/retinal pigment epithelium interface. RESULTS: Patients with AOFVD had a mostly intact photoreceptor layer, a central foveal thickness of 142 +/- 23 microm as well as subretinal deposits. Patients with SD generally had a diffuse degenerative change with a visible reduction in thickness of all intraretinal layers, resulting in a corresponding reduction of central foveal thickness (94 +/- 38 microm) and central loss of photoreceptors (PRs). Comparative central foveal thickness of patients with AOFVD and SD/FF was significantly different (P < .001). Patients with FF had pigment epithelial deposits and paracentral focal photoreceptor loss. CONCLUSIONS: UHR OCT is a clinically feasible tool for examining intraretinal changes, in particular photoreceptor atrophy in macular dystrophies and, therefore, has the potential to be an adequate imaging system for monitoring the course of disease.

Three-dimensional adaptive optics ultrahigh-resolution optical coherence tomography using a liquid crystal spatial light modulator.

A liquid crystal programmable phase modulator (PPM) is used as correcting device in an adaptive optics system for three-dimensional ultrahigh-resolution optical coherence tomography (UHR OCT). The feasibility of the PPM to correct high order aberrations even when using polychromatic light is studied, showing potential for future clinical use. Volumetric UHR OCT of the living retina, obtained with up 25,000A-scans/s and high resolution enables visualization of retinal features that might correspond to groups of terminal bars of photoreceptors at the external limiting membrane.

Three-dimensional ultrahigh-resolution optical coherence tomography of macular diseases.

PURPOSE: To demonstrate a new generation of three-dimensional (3-D) ultrahigh-resolution optical coherence tomography (UHR OCT) technology for visualization of macular diseases. METHODS: One hundred forty eyes with a distinct disease in each of the posterior pole compartments were examined with 3-D UHR OCT. 3-D imaging was performed with a high axial resolution of 3 mum with a compact, commercially available, ultra-broad-bandwidth (160 nm) titanium:sapphire laser at a video rate of up to 25 B-scans/s. Each tomogram consisted of 1024 x 1024 pixels, resulting in 25 megavoxels/s. RESULTS: 3-D UHR OCT offers high-precision 3-D visualization of macular diseases at all structural levels. The UHR modality allows identification of the contour of the hyaloid membrane, tractive forces of epiretinal membranes, and changes within the inner limiting membrane. The system provides quality 3-D images of the topographic dynamics of traction lines from the retinal surface down to the level of the photoreceptor segments. Intraretinal diseases are identified by their specific location in different layers of the neurosensory ultrastructure. Photoreceptor inner and outer segments are clearly delineated in configuration and size, with a characteristic peak in the subfoveal area. The microarchitecture of choroidal neovascularization is distinctly imaged, related leakage can be identified, and the volume can be quantified. CONCLUSIONS: High-speed UHR OCT offers unprecedented, realistic, 3-D imaging of ocular diseases at all epi-, intra- and subretinal levels. A complete 3-D data set of the macular layers allows a comprehensive analysis of focal and diffuse diseases, as well as identification of dynamic pathomechanisms.