Polarization-artifact reduction and accuracy improvement of Jones-matrix polarization-sensitive optical coherence tomography by multi-focus-averaging based multiple scattering reduction.

Polarization-sensitive optical coherence tomography (PS-OCT) is a promising biomedical imaging tool for the differentiation of various tissue properties. However, the presence of multiple-scattering (MS) signals can degrade the quantitative polarization measurement accuracy. We demonstrate a method to reduce MS signals and increase the measurement accuracy of Jones matrix PS-OCT. This method suppresses MS signals by averaging multiple Jones matrix volumes measured using different focal positions. The MS signals are decorrelated among the volumes by focus position modulation and are thus reduced by averaging. However, the single scattering signals are kept consistent among the focus-modulated volumes by computational refocusing. We validated the proposed method using a scattering phantom and a postmortem medaka fish. The results showed reduced artifacts in birefringence and degree-of-polarization uniformity measurements, particularly in deeper regions in the samples. This method offers a practical solution to mitigate MS-induced artifacts in PS-OCT imaging and improves quantitative polarization measurement accuracy.

Multi-focus averaging for multiple scattering suppression in optical coherence tomography.

Multiple scattering is one of the main factors that limits the penetration depth of optical coherence tomography (OCT) in scattering samples. We propose a method termed multi-focus averaging (MFA) to suppress the multiple-scattering signals and improve the image contrast of OCT in deep regions. The MFA method captures multiple OCT volumes with various focal positions and averages them in complex form after correcting the varying defocus through computational refocusing. Because the multiple-scattering takes different trajectories among the different focal position configurations, this averaging suppresses the multiple-scattering signal. Meanwhile, the single-scattering takes a consistent trajectory regardless of the focal position configuration and is not suppressed. Hence, the MFA method improves the ratio between the single-scattering signal and multiple-scattering signal, resulting in an enhancement in the image contrast. A scattering phantom and a postmortem zebrafish were measured to validate the proposed method. The results showed that the contrast of intensity images of both the phantom and zebrafish were improved using the MFA method, such that they were better than the contrast provided by the standard single focus averaging method. The MFA method provides a cost-effective solution for contrast enhancement through multiple-scattering reduction in tissue imaging using OCT systems.

Renal tubular function and morphology revealed in kidney without labeling using three-dimensional dynamic optical coherence tomography.

Renal tubule has distinct metabolic features and functional activity that may be altered during kidney disease. In this paper, we present label-free functional activity imaging of renal tubule in normal and obstructed mouse kidney models using three-dimensional (3D) dynamic optical coherence tomography (OCT) ex vivo. To create an obstructed kidney model, we ligated the ureter of the left kidney for either 7 or 14 days. Two different dynamic OCT (DOCT) methods were implemented to access the slow and fast activity of the renal tubules: a logarithmic intensity variance (LIV) method and a complex-correlation-based method. Three-dimensional DOCT data were acquired with a 1.3 [Formula: see text]m swept-source OCT system and repeating raster scan protocols. In the normal kidney, the renal tubule appeared as a convoluted pipe-like structure in the DOCT projection image. Such pipe-like structures were not observed in the kidneys subjected to obstruction of the ureter for several days. Instead of any anatomical structures, a superficial high dynamics appearance was observed in the perirenal cortex region of the obstructed kidneys. These findings suggest that volumetric LIV can be used as a tool to investigate kidney function during kidney diseases.

Label-free intratissue activity imaging of alveolar organoids with dynamic optical coherence tomography.

An organoid is a three-dimensional (3D) in vitro cell culture emulating human organs. We applied 3D dynamic optical coherence tomography (DOCT) to visualize the intratissue and intracellular activities of human induced pluripotent stem cells (hiPSCs)-derived alveolar organoids in normal and fibrosis models. 3D DOCT data were acquired with an 840-nm spectral domain optical coherence tomography with axial and lateral resolutions of 3.8 µm (in tissue) and 4.9 µm, respectively. The DOCT images were obtained by the logarithmic-intensity-variance (LIV) algorithm, which is sensitive to the signal fluctuation magnitude. The LIV images revealed cystic structures surrounded by high-LIV borders and mesh-like structures with low LIV. The former may be alveoli with a highly dynamics epithelium, while the latter may be fibroblasts. The LIV images also demonstrated the abnormal repair of the alveolar epithelium.

Biospeckle optical coherence tomography in speedy visualizing effects of foliar application of plant growth hormone to Chinese chives leaves.

OBJECTIVE: The aim of this study is to demonstrate the potential of applying the contrast of the speckles obtained as noise in optical coherence tomography (OCT) images to monitor short term activity changes during foliar application of phytohormones to a plant leaf. Plant growth hormone, gibberellic acid (GA3) was sprayed onto the leaf of Chinese chives and after 60 min, OCT images (1 frame: 512 × 2048 pixels) were recorded at ten frames per second for a few tens of seconds. RESULTS: Contrast across the temporal axis was calculated for each pixel of the structural images and biospeckle OCT contrast images were obtained under the conditions of before and after application of GA3 for different concentrations 0, 40, and 100 µM. Application of 40 µM GA3 failed to show any differences in the OCT structural images. However, bOCT contrast image was clearly different. Changes were found to be statistically significant. Although the mechanism for the contrast difference is not clear, it can be said there is a large change across the temporal scale with the application of GA3. Demonstration of OCT utilizing the speckle contrast is believed to have the potential as a promising tool in plant physiology.

Developmental and morphological studies in Japanese medaka with ultra-high resolution optical coherence tomography.

We propose ultra-high resolution optical coherence tomography to study the morphological development of internal organs in medaka fish in the post-embryonic stages at micrometer resolution. Different stages of Japanese medaka were imaged after hatching in vivo with an axial resolution of 2.8 µm in tissue. Various morphological structures and organs identified in the OCT images were then compared with the histology. Due to the medaka's close resemblance to vertebrates, including humans, these morphological features play an important role in morphogenesis and can be used to study diseases that also occur in humans.

Comparison of three-dimensional optical coherence tomography and combining a rotating Scheimpflug camera with a Placido topography system for forme fruste keratoconus diagnosis.

OBJECTIVE: To evaluate the ability of parameters measured by three-dimensional (3D) corneal and anterior segment optical coherence tomography (CAS-OCT) and a rotating Scheimpflug camera combined with a Placido topography system (Scheimpflug camera with topography) to discriminate between normal eyes and forme fruste keratoconus. METHODS: Forty-eight eyes of 48 patients with keratoconus, 25 eyes of 25 patients with forme fruste keratoconus and 128 eyes of 128 normal subjects were evaluated. Anterior and posterior keratometric parameters (steep K, flat K, average K), elevation, topographic parameters, regular and irregular astigmatism (spherical, asymmetry, regular and higher-order astigmatism) and five pachymetric parameters (minimum, minimum-median, inferior-superior, inferotemporal-superonasal, vertical thinnest location of the cornea) were measured using 3D CAS-OCT and a Scheimpflug camera with topography. The area under the receiver operating curve (AUROC) was calculated to assess the discrimination ability. Compatibility and repeatability of both devices were evaluated. RESULTS: Posterior surface elevation showed higher AUROC values in discrimination analysis of forme fruste keratoconus using both devices. Both instruments showed significant linear correlations (p<0.05, Pearson's correlation coefficient) and good repeatability (ICCs: 0.885-0.999) for normal and forme fruste keratoconus. CONCLUSIONS: Posterior elevation was the best discrimination parameter for forme fruste keratoconus. Both instruments presented good correlation and repeatability for this condition.

Advanced multi-contrast Jones matrix optical coherence tomography for Doppler and polarization sensitive imaging.

An advanced version of Jones matrix optical coherence tomography (JMT) is demonstrated for Doppler and polarization sensitive imaging of the posterior eye. JMT is capable of providing localized flow tomography by Doppler detection and investigating the birefringence property of tissue through a three-dimensional (3-D) Jones matrix measurement. Owing to an incident polarization multiplexing scheme based on passive optical components, this system is stable, safe in a clinical environment, and cost effective. Since the properties of this version of JMT provide intrinsic compensation for system imperfection, the system is easy to calibrate. Compared with the previous version of JMT, this advanced JMT achieves a sufficiently long depth measurement range for clinical cases of posterior eye disease. Furthermore, a fine spectral shift compensation method based on the cross-correlation of calibration signals was devised for stabilizing the phase of OCT, which enables a high sensitivity Doppler OCT measurement. In addition, a new theory of JMT which integrates the Jones matrix measurement, Doppler measurement, and scattering measurement is presented. This theory enables a sensitivity-enhanced scattering OCT and high-sensitivity Doppler OCT. These new features enable the application of this system to clinical cases. A healthy subject and a geographic atrophy patient were measured in vivo, and simultaneous imaging of choroidal vasculature and birefringence structures are demonstrated.

Keratoconus diagnosis using anterior segment polarization-sensitive optical coherence tomography.

PURPOSE: To investigate the tissue properties of keratoconic and normal corneas in vivo by using polarization-sensitive optical coherence tomography (PS-OCT), and to evaluate early keratoconus by the area under the receiver operating characteristic curve (AUROC) and Mahalanobis distance analysis. METHODS: Thirty-one eyes of 20 patients with keratoconus, 7 eyes of 7 patients with keratoconus suspect, and 25 eyes of 25 normal subjects were investigated by PS-OCT and corneal and anterior segment (CAS)-OCT. Average of en face phase retardation of the posterior surface of the cornea, curvature, videokeratographic parameters, regular and irregular astigmatism, pachymetry map, and elevation were measured. The AUROC of each parameter was calculated to evaluate the diagnostic power to detect keratoconus and keratoconus suspect. RESULTS: While in normal controls, the center of the en face phase retardation map showed low and homogeneous birefringence, in keratoconic corneas the birefringence increased with disease severity. Some keratoconus suspects had apparent high birefringence values without displaying morphologic signs of keratoconus in the anterior surface. The phase retardation values for normal, keratoconus suspect, and keratoconus subjects were, respectively, 0.20 ± 0.06, 0.35 ± 0.06, and 0.50 ± 0.14 radians in 3 mm diameter and 0.29 ± 0.22, 0.33 ± 0.13, and 0.74 ± 0.45 radians in 6 mm diameter. Outer morphological parameters such as curvature, videokeratographic parameters, regular and irregular astigmatism, and elevation showed high AUROCs for discriminating keratoconus from normal controls. On the other hand, using Mahalanobis distance, the AUROC of phase retardation, which represents microstructural properties of tissue, showed high value (0.989-1.000) for discriminating between keratoconus suspects and normal controls. CONCLUSIONS: Average of en face phase retardation of the posterior surface of the cornea was increased in keratoconus patients due to changes in the lamellar structure of collagen fibers. Phase retardation was sensitive with regard to discriminating keratoconus suspect and might be useful for detecting very early or even subclinical keratoconus.

Optical rheology of porcine sclera by birefringence imaging.

PURPOSE: To investigate a relationship between birefringence and elasticity of porcine sclera ex vivo using polarization-sensitive optical coherence tomography (PS-OCT). METHODS: Elastic parameters and birefringence of 19 porcine eyes were measured. Four pieces of scleral strips which were parallel to the limbus, with a width of 4 mm, were dissected from the optic nerve head to the temporal side of each porcine eye. Birefringence of the sclera was measured with a prototype PS-OCT. The strain and force were measured with a uniaxial material tester as the sample was stretched with a speed of 1.8 mm/min after preconditioning. A derivative of the exponentially-fitted stress-strain curve at 0% strain was extracted as the tangent modulus. Power of exponential stress-strain function was also extracted from the fitting. To consider a net stiffness of sclera, structural stiffness was calculated as a product of tangent modulus and thickness. Correlations between birefringence and these elastic parameters were examined. RESULTS: Statistically significant correlations between birefringence and all of the elastic parameters were found at 2 central positions. Structural stiffness and power of exponential stress-strain function were correlated with birefringence at the position near the optic nerve head. No correlation was found at the position near the equator. CONCLUSIONS: The evidence of correlations between birefringence and elasticity of sclera tested uniaxially was shown for the first time. This work may become a basis for in vivo measurement of scleral biomechanics using PS-OCT.

Passive component based multifunctional Jones matrix swept source optical coherence tomography for Doppler and polarization imaging.

We present a fiber based multifunctional Jones matrix swept source optical coherence tomography (SS-OCT) system for Doppler and polarization imaging. Jones matrix measurement without using active components such as electro-optic modulators is realized by incident polarization multiplexing based on independent delay of two orthogonal polarization states and polarization diversity detection. In addition to polarization sensitivity, this system measures Doppler flow without extra hardware for phase stabilized SS-OCT detection. An eighth-wave plate was measured to demonstrate the polarization detection accuracy. The optic nerve head of a retina was measured in vivo. Detailed vasculature and birefringent structures were investigated simultaneously.

In vivo evaluation of human skin anisotropy by polarization-sensitive optical coherence tomography.

We performed an in vivo three-dimensional analysis of anisotropic changes in the dermal birefringence of mechanically deformed human skin using polarization-sensitive optical coherence tomography (PS-OCT). The papillary-dermal birefringence of the forehead increased significantly when the skin was shrunk parallel to the body axis, and decreased significantly when the skin was shrunk perpendicular to the body axis. En-face images of the papillary-dermal birefringence revealed variations among individual subjects, and that both shrinking parallel to and stretching in perpendicular to the body axis promoted the formation of macro rope-like birefringent domains. We found that PS-OCT is useful for understanding anisotropic properties of collagen structure in the skin.

Monte-Carlo-based phase retardation estimator for polarization sensitive optical coherence tomography.

A Monte-Carlo-based phase retardation estimator is developed to correct the systematic error in phase retardation measurement by polarization sensitive optical coherence tomography (PS-OCT). Recent research has revealed that the phase retardation measured by PS-OCT has a distribution that is neither symmetric nor centered at the true value. Hence, a standard mean estimator gives us erroneous estimations of phase retardation, and it degrades the performance of PS-OCT for quantitative assessment. In this paper, the noise property in phase retardation is investigated in detail by Monte-Carlo simulation and experiments. A distribution transform function is designed to eliminate the systematic error by using the result of the Monte-Carlo simulation. This distribution transformation is followed by a mean estimator. This process provides a significantly better estimation of phase retardation than a standard mean estimator. This method is validated both by numerical simulations and experiments. The application of this method to in vitro and in vivo biological samples is also demonstrated.

Birefringence measurement of cornea and anterior segment by office-based polarization-sensitive optical coherence tomography.

We present a case series of cornea and anterior segment disorders investigated by an office-based polarization-sensitive optical coherence tomography (PS-OCT). Blebs of glaucoma patients treated by trabeculectomy, and corneas of keratoconus and keratoplasty patients were measured by PS-OCT. Birefringence formations in trabeculectomy bleb were measured in 1 control eye and 3 eyes of trabeculectomy model rabbits. Polarization insensitive scattering OCT and the depth-resolved birefringence were measured simultaneously by PS-OCT. Abnormal birefringence was observed in keratoconus cases with advanced thinning and with a rupture of Descemet's membrane. The graft-host interface of the keratoplasty case showed abnormal birefringence. The appearance of abnormal birefringence in the cornea was likely to be an indication of cross-linking of collagen fibrils. The measurement of rabbit showed abnormal birefringence in the scarring eyes. Wide regions of strong birefringence were observed in the eyes of trabeculectomy patients who had high intraocular pressure. Visualization of scarring in bleb by PS-OCT may be useful for the planning of secondary surgery. PS-OCT showed promising for the study and diagnosis diseases related to abnormal fibrous tissues of the cornea and anterior eye segment.

Multimodal analysis of pearls and pearl treatments by using optical coherence tomography and fluorescence spectroscopy.

We present an integrated optical system that consists of optical coherence tomography (OCT) and laser-induced fluorescence (LIF) spectroscopy for multimodal analysis of pearls and pearl treatments. The OCT source and the LIF excitation beams were aligned together to illuminate the same spot of a pearl fixed on the sample stage that was under rotation. As a result, both OCT images and LIF spectra of the pearls were detected at the same time and also at the same place. For OCT, a 1310 nm-centered swept laser source was used. For LIF, a 405 nm laser diode was used and a lensed multimode fiber was utilized as a fluorescence probe. The tomographic investigation on the internal structure of a pearl allowed us to evaluate and categorize the pearl nondestructively as was previously reported. In addition, the measurements of fluorescence spectrum and its decaying rate helped to determine the species of mother oyster. The proposed multimodal analysis made it possible to classify the pearls and also to disclose the treatments made on the pearls.

Full-range polarization-sensitive swept-source optical coherence tomography by simultaneous transversal and spectral modulation.

Polarization-sensitive swept-source optical coherence tomography (PS-SS-OCT) is used to measure three-dimensional phase-retardation images of birefringent biological tissue in vivo. PS-SS-OCT with continuous source polarization modulation is used to multiplex the incident states of polarization in the signal frequency of each A-scan. Although it offers the advantage of measurement speed that is as high as that of standard SS-OCT, its disadvantage is low axial measurement range. To overcome this drawback, we employed the B-M-mode scan (BM-scan) method, which removes complex conjugate ambiguity by applying phase modulation along the transversal scanning direction. Since polarization modulation and BM-scan are applied in different scanning directions, these methods can be combined to make the optimum use of both full range and polarization-sensitive imaging. Phase fluctuations that cause measurement failure were numerically canceled before demodulating the B-scan oriented modulation. After removing complex conjugate artifacts, the axial measurement range was 5.35 mm, and the signal-to-conjugate ratio was 40.5 dB. We demonstrated retinal imaging using the PS-SS-OCT system with a frequency-swept laser at a center wavelength of 1064 nm and an axial resolution of 11.4 microm in tissue. Full-range polarization-sensitive retinal images showed characteristic birefringence of fibrous tissues such as retinal nerve fiber, sclera, and lamina cribrosa.

Visualization of phase retardation of deep posterior eye by polarization-sensitive swept-source optical coherence tomography with 1-microm probe.

Polarization-sensitive optical coherence tomography (PS-OCT) can measure cross-sectional and volumetric images of birefringence in fibrous tissues that provides additional contrast to the intensity images. In this study, we develop polarization-sensitive swept-source OCT (PS-SS-OCT) at 1 microm for deep penetration of the sclera and lamina cribrosa in the posterior part of human eyes. A calibration method for polarization mode dispersion of a circulator, which is employed to conserve the optical power of the interferometer and achieve system sensitivity sufficient for retinal imaging is demonstrated. The A-scan rate, the axial resolution, and the sensitivity of the PS-SS-OCT are 28,000 Hz, 11.0 microm, 94.2 dB, respectively. The posterior part of the eyes of a healthy male subject are measured in vivo. Phase-retardation images show birefringence of deep sclera and lamina cribrosa and enhance the contrast which is not visible in the intensity images. In addition, unlike conventional OCT, our PS-SS-OCT showed polarization-insensitive intensity images, in which an artifact created by the birefringence of sclera has been successfully eliminated.