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.

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.

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.

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.

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.

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.

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.

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.

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.

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

Assessment of central visual function in Stargardt's disease/fundus flavimaculatus with ultrahigh-resolution optical coherence tomography.

PURPOSE: To assess photoreceptor morphology in patients with Stargardt's disease and fundus flavimaculatus using ultrahigh-resolution optical coherence tomography (UHR-OCT) and correlate it with visual acuity (VA). METHODS: This was a prospective observational case series. Fourteen patients with Stargardt's disease (nine women, five men; average age, 39 years; range, 27-53) were examined. A clinically viable UHR-OCT system employing a new, compact titanium sapphire laser was used, enabling a 3-microm axial resolution in the retina. All patients received a full ophthalmic examination, including fluorescein angiography. Outcome was judged by central transverse photoreceptor loss, central foveal thickness, VA, central atrophy according to fluorescein angiography, and fundus autofluorescence. RESULTS: UHR-OCT was capable of visualizing and quantifying regions of central transverse photoreceptor (PR) loss. All Stargardt patients with central atrophy had a complete loss of the central photoreceptor layer in the foveal region (mean transverse photoreceptor loss, 4390 +/- 2270 microm; range, 530-9240 microm). Patients without clinically evident central atrophy had an intact photoreceptor layer centrally, but had small, focal parafoveal defects. A correlation was detected between VA and transverse PR loss (Spearman rho=-0.60, P=0.03), which was confirmed on logistic regression analysis (R2=0.49, P=0.0001). Central foveal thickness was reduced in patients with Stargardt's disease (85 +/- 40 microm; range, 58-280 microm). The correlation was statistically significant with VA (Spearman rho=0.43, P=0.04), but not with transverse PR loss (Spearman rho=-0.23, P >>0.05). Linear regression analysis showed a statistically significant association of central foveal thickness with VA (R2=0.51, P=0.0001), but not with transverse PR loss (P >>0.05). The extent of atrophy seen in fluorescein angiography correlated with VA and transverse PR loss (Spearman rho=-0.51, P=0.007; Spearman rho=0.77, P=0.0001). Similar correlations were found with the maximum transverse diameter of fundus autofluorescence (Spearman rho=-0.72, P=0.008; Spearman rho=0.77, P=0.003). CONCLUSIONS: Ultrahigh-resolution OCT demonstrates excellent visualization of intraretinal morphology and enables quantification of the photoreceptor layer. Thus, for the first time, an in vivo visualization and quantification of transverse, central photoreceptor loss and correlation with visual function is possible. Lower VA corresponds to a greater transverse photoreceptor loss, which also correlates with the extent of changes seen in fluorescein angiography and in fundus autofluorescence. Furthermore, reduced retinal thickness (i.e., atrophy of retinal layers) does not correlate with the transverse extent of PR loss. Thus, it seems that although there may be progressive atrophy of intraretinal layers, an intact photoreceptor layer leads to better VA. UHR-OCT may present a viable alternative to the assessment of central visual function, due to the easy, objective, and noninvasive data acquisition. Therefore, UHR-OCT could be of future use in judging patients' prognoses in Stargardt's disease.

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.

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.

Imaging ex vivo healthy and pathological human brain tissue with ultra-high-resolution optical coherence tomography.

The ability of ultra-high-resolution optical coherence tomography (UHR OCT) to discriminate between healthy and pathological human brain tissue is examined by imaging ex vivo tissue morphology of various brain biopsies. Micrometer-scale OCT resolution (0.9x2 microm, axialxlateral) is achieved in biological tissue by interfacing a state-of-the-art Ti:Al2O3 laser (lambda(c)=800 nm, delta lambda=260 nm, and P(out)=120 mW exfiber) to a free-space OCT system utilizing dynamic focusing. UHR OCT images are acquired from both healthy brain tissue and various types of brain tumors including fibrous, athypical, and transitional meningioma and ganglioglioma. A comparison of the tomograms with standard hematoxylin and eosin (H&E) stained histological sections of the imaged biopsies demonstrates the ability of UHR OCT to visualize and identify morphological features such as microcalcifications (>20 microm), enlarged nuclei of tumor cells (approximately 8 to 15 microm), small cysts, and blood vessels, which are characteristic of neuropathologies and normally absent in healthy brain tissue.

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.

Intraocular lens-capsular bag imaging with ultrahigh-resolution optical coherence tomography Pseudophakic human autopsy eyes.

PURPOSE: To compare in vitro ultrahigh-resolution optical coherence tomography (UHR OCT) cross-sectional images of the pseudophakic human autopsy eye with histology to evaluate the potential of this imaging technique for enhanced visualization of the anterior segment, especially the capsular bag, intraocular lens (IOL), and posterior capsule opacification (PCO) formation after cataract surgery. SETTING: Department of Medical Physics and Department of Ophthalmology, University of Vienna, Vienna, Austria, and Department of Oral and Maxillofacial Surgery Institute of Dentistry, University of Turku, Turku, Finland. METHODS: Ultrahigh-resolution OCT images were acquired from 7 pseudophakic human autopsy eyes using 1.4 microm axial x 3.0 microm transverse resolution. The axial resolution with UHR OCT is 1.4 microm compared to 10.0 microm with the commercially available OCT. Plastic-embedded histologic sections were obtained in precise alignment with the OCT tomograms. RESULTS: Ultrahigh-resolution OCT cross-sectional tomograms corresponded to the histologic sections. With the wavelength used (800 nm), the anterior and posterior capsules, area of lens epithelial cell growth and extracellular matrix proliferation, and IOL could be clearly visualized. The extent of capsular bag adhesion to the IOL could be detected, as well as the amount of PCO formation. CONCLUSIONS: The improved resolution makes UHR OCT a powerful tool in anterior segment imaging and evaluation of the capacity of IOL materials and models to induce capsular bag adhesion. Ultrahigh-resolution OCT may also help in determining the area of origin of PCO after cataract surgery.

Histologic correlation of pig retina radial stratification with ultrahigh-resolution optical coherence tomography.

PURPOSE: To compare ultrahigh-resolution optical coherence tomography (OCT) cross-sectional images of the pig retina with histology, to evaluate the potential of ultrahigh-resolution OCT for enhanced visualization of intra- and subretinal structures. METHODS: Ultrahigh-resolution OCT images were acquired with 1.4- micro m axial x 3- micro m transverse resolution from in vitro posterior eyecup preparations of the domestic pig. Frozen sections were obtained in precise alignment with OCT tomograms, by using major blood vessels as orientation markers and were counterstained with cresyl violet or unstained and examined by differential interference contrast microscopy. Micrographs from histologic sections were linearly scaled to correct for tissue shrinkage and compared with OCT tomograms. RESULTS: In the proximal retina, ultrahigh-resolution OCT signal bands directly corresponded to the main retinal layers. For the wavelength region used ( approximately 800 nm), axodendritic layers (nerve fiber layer, inner and outer plexiform layers) were more reflective than cell body layers (ganglion cell layer, inner nuclear layer, outer nuclear layer). In the distal retina, substructures of the photoreceptor layer such as the interface between inner and outer segments were visualized, and the retinal pigment epithelium, the choriocapillaris, and superficial choroid layers were resolved. In addition, the time sequence of a retinal detachment event was monitored by ultrahigh-resolution OCT. CONCLUSIONS: In vitro ophthalmic ultrahigh-resolution OCT imaging reveals retinal morphology with unprecedented detail. The specific assignment of OCT signal patterns to retinal substructures provides a basis for improved interpretation of in vivo ophthalmic OCT tomograms of high clinical relevance.

Imaging of polarization properties of human retina in vivo with phase resolved transversal PS-OCT.

Recently, we developed a phase resolved polarization sensitive OCT system based on transversal scanning. This system was now improved and adapted for retinal imaging in vivo. We accelerated the image acquisition speed by a factor of 10 and adapted the system for light sources emitting at 820nm. The improved instrument records 1000 transversal lines per second. Two different scanning modes enable either the acquisition of high resolution B-scan images containing 1600x500 pixels in 500ms or the recording of 3D data sets by C-scan mode imaging. This allows acquiring a 3D-data set containing 1000x100x100 pixels in 10 seconds. We present polarization sensitive B-scan images and to the best of our knowledge, the first 3D-data sets of retardation and fast axis orientation of fovea and optic nerve head region in vivo. The polarizing and birefringence properties of different retinal layers: retinal pigment epithelium, Henle's fiber layer, and retinal nerve fiber layer are studied.

Enhanced visualization of macular pathology with the use of ultrahigh-resolution optical coherence tomography.

OBJECTIVES: To demonstrate a new generation of ophthalmic optical coherence tomography (OCT) technology with unprecedented axial resolution for enhanced imaging of intraretinal microstructures and to investigate its clinical feasibility to visualize intraretinal morphology of macular pathology. METHODS: A clinically viable ultrahigh-resolution ophthalmic OCT system was developed and used in clinical imaging for the first time. Fifty-six eyes of 40 selected patients with different macular diseases including macular hole, macular edema, age-related macular degeneration, central serous chorioretinopathy, epiretinal membranes, and detachment of pigment epithelium and sensory retina were included. OUTCOME MEASURES: Ultrahigh-resolution tomograms visualizing intraretinal morphologic features in different retinal diseases. RESULTS: An axial image resolution of approximately 3 micro m was achieved in the eyes examined, nearly 2 orders of magnitude better than conventional ophthalmic ultrasound. Ultrahigh-resolution OCT images provided additional diagnostically important information on intraretinal morphologic features that could not have been obtained by standard techniques. CONCLUSIONS: Ultrahigh-resolution ophthalmic OCT enables unprecedented visualization of intraretinal morphologic features and therefore has the potential to contribute to a better understanding of ocular pathogenesis, as well as to enhance the sensitivity and specificity for early ophthalmic diagnosis and to monitor the efficacy of therapy. This study establishes a baseline for the interpretation of ultrahigh-resolution ophthalmic OCT imaging of macular diseases.