In vivo functional microangiography by visible-light optical coherence tomography

Although hemoglobin oxygen saturation (sO₂) in the microvasculature is an essential physiological parameter of local tissue functions, non-invasive measurement of microvascular sO₂ is still challenging. Here, we demonstrated that visible-light optical coherence tomography (vis-OCT) can simultaneously provide three-dimensional anatomical tissue morphology, visualize microvasculature at the capillary level, and measure sO₂ from the microvasculature in vivo. We utilized speckle contrast caused by the moving blood cells to enhance microvascular imaging. We applied a series of short-time inverse Fourier transforms to obtain the spectroscopic profile of blood optical attenuation, from which we quantified sO₂. We validated the sO₂ measurement in mouse ears in vivo through hypoxia and hyperoxia challenges. We further demonstrated that vis-OCT can continuously monitor dynamic changes of microvascular sO_2.OCIS codes: (170.4500) Optical coherence tomography, (300.1030) Absorption     

Quantitative comparison of analysis methods for spectroscopic optical coherence tomography

Spectroscopic optical coherence tomography (sOCT) enables the mapping of chromophore concentrations and image contrast enhancement in tissue. Acquisition of depth resolved spectra by sOCT requires analysis methods with optimal spectral/spatial resolution and spectral recovery. In this article, we quantitatively compare the available methods, i.e. the short time Fourier transform (STFT), wavelet transforms, the Wigner-Ville distribution and the dual window method through simulations in tissue-like media. We conclude that all methods suffer from the trade-off in spectral/spatial resolution, and that the STFT is the optimal method for the specific application of the localized quantification of hemoglobin concentration and oxygen saturation.OCIS codes: (030.1640) Coherence, (070.4790) Spectrum analysis, (160.4760) Optical properties, (170.6510) Spectroscopy, tissue diagnostics     

Depth resolved detection of lipid using spectroscopic optical coherence tomography

Optical frequency domain imaging (OFDI) can identify key components related to plaque vulnerability but can suffer from artifacts that could prevent accurate identification of lipid rich regions. In this paper, we present a model of depth resolved spectral analysis of OFDI data for improved detection of lipid. A quadratic Discriminant analysis model was developed based on phantom compositions known chemical mixtures and applied to a tissue phantom of a lipid-rich plaque. We demonstrate that a combined spectral and attenuation model can be used to predict the presence of lipid in OFDI images.OCIS codes: (110.4500) Optical coherence tomography, (170.1580) Chemometrics, (170.6510) Spectroscopy, tissue diagnostics     

Multispectral nanoparticle contrast agents for true-color spectroscopic optical coherence tomography

We have recently developed a novel dual window scheme for processing spectroscopic OCT images to provide spatially resolved true color imaging of chromophores in scattering samples. Here we apply this method to measure the extinction spectra of plasmonic nanoparticles at various concentrations for potential in vivo applications. We experimentally demonstrate sub-nanomolar sensitivity in the measurement of nanoparticle concentrations, and show that colorimetric imaging with multiple species of nanoparticles produces enhanced contrast for spectroscopic OCT in both tissue phantom and cell studies.     

Human retinal imaging using visible-light optical coherence tomography guided by scanning laser ophthalmoscopy

We achieved human retinal imaging using visible-light optical coherence tomography (vis-OCT) guided by an integrated scanning laser ophthalmoscopy (SLO). We adapted a spectral domain OCT configuration and used a supercontinuum laser as the illumating source. The center wavelength was 564 nm and the bandwidth was 115 nm, which provided a 0.97 µm axial resolution measured in air. We characterized the sensitivity to be 86 dB with 226 µW incidence power on the pupil. We also integrated an SLO that shared the same optical path of the vis-OCT sample arm for alignment purposes. We demonstrated the retinal imaging from both systems centered at the fovea and optic nerve head with 20° × 20° and 10° × 10° field of view. We observed similar anatomical structures in vis-OCT and NIR-OCT. The contrast appeared different from vis-OCT to NIR-OCT, including slightly weaker signal from intra-retinal layers, and increased visibility and contrast of anatomical layers in the outer retina.OCIS codes: (110.4190) Multiple imaging, (170.0110) Imaging systems, (170.4470) Ophthalmology, (170.4500) Optical coherence tomography     

Comparison of different metrics for analysis and visualization in spectroscopic optical coherence tomography

Spectroscopic Optical Coherence Tomography (S-OCT) extracts depth resolved spectra that are inherently available from OCT signals. The back scattered spectra contain useful functional information regarding the sample, since the light is altered by wavelength dependent absorption and scattering caused by chromophores and structures of the sample. Two aspects dominate the performance of S-OCT: (1) the spectral analysis processing method used to obtain the spatially-resolved spectroscopic information and (2) the metrics used to visualize and interpret relevant sample features. In this work, we focus on the second aspect, where we will compare established and novel metrics for S-OCT. These concepts include the adaptation of methods known from multispectral imaging and modern signal processing approaches such as pattern recognition. To compare the performance of the metrics in a quantitative manner, we use phantoms with microsphere scatterers of different sizes that are below the system’s resolution and therefore cannot be differentiated using intensity based OCT images. We show that the analysis of the spectral features can clearly separate areas with different scattering properties in multi-layer phantoms. Finally, we demonstrate the performance of our approach for contrast enhancement in bovine articular cartilage.OCIS codes: (170.4500) Optical coherence tomography, (300.0300) Spectroscopy, (290.5850) Scattering, particles, (180.0180) Microscopy, (170.3880) Medical and biological imaging     

Multiple forward scattering reduces the measured scattering coefficient of whole blood in visible-light optical coherence tomography

The optical properties of blood encode oxygen-dependent information. Noninvasive optical detection of these properties is increasingly desirable to extract biomarkers for tissue health. Recently, visible-light optical coherence tomography (vis-OCT) demonstrated retinal oxygen saturation (sO₂) measurements by inversely measuring the oxygen-dependent absorption and scattering coefficients of whole blood. However, vis-OCT may be sensitive to optical scattering properties of whole blood, different from those reported in the literature. Incorrect assumptions of such properties can add additional uncertainties or biases to vis-OCT’s sO₂ model. This work investigates whole blood’s scattering coefficient measured by vis-OCT. Using Monte Carlo simulation of a retinal vessel, we determined that vis-OCT almost exclusively detects multiple-scattered photons in whole blood. Meanwhile, photons mostly forward scatter in whole blood within the visible spectral range, allowing photons to maintain ballistic paths and penetrate deeply, leading to a reduction in the measured scattering coefficient. We defined a scattering scaling factor (SSF) to account for such a reduction and found that SSF varied with measurement conditions, such as numerical aperture, depth resolution, and depth selection. We further experimentally validated SSF in ex vivo blood phantoms with pre-set sO₂ levels and in the human retina, both of which agreed well with our simulation.     

Assessing hemoglobin concentration using spectroscopic optical coherence tomography for feasibility of tissue diagnostics

Hemoglobin (Hb) concentration and oxygen saturation levels are important biomarkers for various diseases, including cancer. Here, we investigate the ability to measure these parameters for tissue using spectroscopic optical coherence tomography (SOCT). A parallel frequency domain OCT system is used with detection spanning the visible region of the spectrum (450 nm to 700 nm). Oxygenated and deoxygenated Hb absorbing phantoms are analyzed. The results show that Hb concentrations as low as 1.2 g/L at 1 mm can be retrieved indicating that both normal and cancerous tissue measurements may be obtained. However, measurement of oxygen saturation levels may not be achieved with this approach.     

Evaluation of burn severity in vivo in a mouse model using spectroscopic optical coherence tomography

Clinical management of burn injuries depends upon an accurate assessment of the depth of the wound. Current diagnostic methods rely primarily on subjective visual inspection, which can produce variable results. In this study, spectroscopic optical coherence tomography was used to objectively evaluate burn injuries in vivo in a mouse model. Significant spectral differences were observed and correlated with the depth of the injury as determined by histopathology. The relevance of these results to clinical burn management in human tissues is discussed.OCIS codes: (170.6510) Spectroscopy, tissue diagnostics; (110.4500) Optical coherence tomography     

Volumetric directional optical coherence tomography

Photoreceptor loss and resultant thinning of the outer nuclear layer (ONL) is an important pathological feature of retinal degenerations and may serve as a useful imaging biomarker for age-related macular degeneration. However, the demarcation between the ONL and the adjacent Henle’s fiber layer (HFL) is difficult to visualize with standard optical coherence tomography (OCT). A dedicated OCT system that can precisely control and continuously and synchronously update the imaging beam entry points during scanning has not been realized yet. In this paper, we introduce a novel imaging technology, Volumetric Directional OCT (VD-OCT), which can dynamically adjust the incident beam on the pupil without manual adjustment during a volumetric OCT scan. We also implement a customized spoke-circular scanning pattern to observe the appearance of HFL with sufficient optical contrast in continuous cross-sectional scans through the entire volume. The application of VD-OCT for retinal imaging to exploit directional reflectivity properties of tissue layers has the potential to allow for early identification of retinal diseases.     

Quantitative comparison of analysis methods for spectroscopic optical coherence tomography: reply to comment

We reply to the comment by Kraszewski et al on “Quantitative comparison of analysis methods for spectroscopic optical coherence tomography.” We present additional simulations evaluating the proposed window function. We conclude that our simulations show good qualitative agreement with the results of Kraszewski, in support of their conclusion that SOCT optimization should include window shape, next to choice of window size and analysis algorithm.OCIS codes: (030.1640) Coherence, (070.4790) Spectrum analysis, (160.4760) Optical properties, (170.6510) Spectroscopy, tissue diagnostics     

Approximate image synthesis in optical coherence tomography

Full-wave models of OCT image formation, which are based on Maxwell’s equations, are highly realistic. However, such models incur a high computational cost, particularly when modelling sample volumes consistent with those encountered in practice. Here, we present an approximate means of synthesizing volumetric image formation to reduce this computational burden. Instead of performing a full-wave scattered light calculation for each A-scan, we perform a full-wave scattered light calculation for a normally incident plane wave only. We use the angular spectrum field representation to implement beam focussing and scanning, making use of an assumption similar to the tilt optical memory effect, to approximately synthesize volumetric data sets. Our approach leads to an order of magnitude reduction in the computation time required to simulate typical B-scans. We evaluate this method by comparing rigorously and approximately evaluated point spread functions and images of highly scattering structured samples for a typical OCT system. Our approach also reveals new insights into image formation in OCT.     

Conical scan polarization-sensitive optical coherence tomography

We report on a new articular cartilage imaging technique with potential for clinical arthroscopic use, by supplementing the variable-incidence-angle polarization-sensitive optical coherence tomography method previously developed by us with a conical beam scan protocol. The technique is validated on bovine tendon by comparing experimental data with simulated data generated using the extended Jones matrix calculus. A unique capability of this new optical technique is that it can locate the “brushing direction” of collagen fibers in articular cartilage, which is structural information that extends beyond established methods such as split-line photography or birefringent fast-axis measurement in that it is uniquely defined over the full azimuthal-angle range of (-π, + π). The mapping of this direction over the cartilage surface may offer insights into the optimal design of tissue-engineering scaffolds for cartilage repair.OCIS codes: (170.4500) Optical coherence tomography, (170.3880) Medical and biological imaging, (170.3890) Medical optics instrumentation, (260.1440) Birefringence, (260.5430) Polarization     

Photothermal optical lock-in optical coherence tomography for in vivo imaging

Photothermal OCT (PTOCT) provides high sensitivity to molecular targets in tissue, and occupies a spatial imaging regime that is attractive for small animal imaging. However, current implementations of PTOCT require extensive temporal sampling, resulting in slow frame rates and a large data burden that limit its in vivo utility. To address these limitations, we have implemented optical lock-in techniques for photothermal optical lock-in OCT (poli-OCT), and demonstrated the in vivo imaging capabilities of this approach. The poli-OCT signal was assessed in tissue-mimicking phantoms containing indocyanine green (ICG), an FDA approved small molecule that has not been previously imaged in vivo with PTOCT. Then, the effects of in vivo blood flow and motion artifact were assessed and attenuated, and in vivo poli-OCT was demonstrated with both ICG and gold nanorods as contrast agents. Experiments revealed that poli-OCT signals agreed with optical lock-in theory and the bio-heat equation, and the system exhibited shot noise limited performance. In phantoms containing biologically relevant concentrations of ICG (1 µg/ml), the poli-OCT signal was significantly greater than control phantoms (p<0.05), demonstrating sensitivity to small molecules. Finally, in vivo poli-OCT of ICG identified the lymphatic vessels in a mouse ear, and also identified low concentrations (200 pM) of gold nanorods in subcutaneous injections at frame rates ten times faster than previously reported. This work illustrates that future in vivo molecular imaging studies could benefit from the improved acquisition and analysis times enabled by poli-OCT.OCIS codes: (110.4500) Optical coherence tomography, (350.5340) Photothermal effects     

光学相干层析成像技术在宫颈疾病中的应用

目的:探讨光学相干层析成像技术(optical coherence tomography,OCT)在宫颈疾病中的应用价值。方法:郑州大学第三附属医院131例宫颈液基细胞学检查(thinprep cytologic test,TCT)结果阳性或人乳头瘤病毒(human papillomavirus,HPV)高危型阳性的患者被纳入研究。被研究者均行OCT及阴道镜配合下宫颈组织活检术,以宫颈组织病理结果作为诊断“金标准”,评价3种方法的灵敏度、特异度及Kappa值。结果:从OCT图像中可以清楚地区分正常宫颈、炎症、低度鳞状上皮内病变(low-grade squamous intraepithelial lesions,LSIL)、高度鳞状上皮内病变(high-grade squamous intraepithelial lesions,HSIL)和浸润性宫颈癌5种型别,其与相应的组织学切片的检查结果匹配良好。HPV灵敏度最高为98.25%,其次是OCT为84.21%,两者差异无统计学意义(P>0.05),均高于TCT检查(P<0.05);TCT特异度为35.14%,HPV为13.51%,与OCT(81.08%)相比差异均存在统计学意义(P<0.05);HPV,TCT及OCT三者的Kappa值分别为0.104,0.017和0.646。结论:OCT作为一种无创、高分辨率、实时快速成像技术,可以与病理切片相媲美,且具有较高的灵敏度与特异度。     

Origins of subdiffractional contrast in optical coherence tomography

We demonstrate that OCT images quantify subdiffractional tissue structure. Optical coherence tomography (OCT) measures stratified tissue morphology with spatial resolution limited by the temporal coherence length. Spectroscopic OCT processing, on the other hand, has enabled nanoscale sensitive analysis, presenting an unexplored question: how does subdiffractional information get folded into the OCT image and how does one best analyze to allow for unambiguous quantification of ultrastructure? We first develop an FDTD simulation to model spectral domain OCT with nanometer resolution. Using this, we validate an analytical relationship between the sample statistics through the power spectral density (PSD) of refractive index fluctuations and three measurable quantities (image mean, image variance, and spectral slope), and have found that each probes different aspects of the PSD (amplitude, integral and slope, respectively). Finally, we found that only the spectral slope, quantifying mass scaling, is monotonic with the sample autocorrelation shape.     

Tissue thickness calculation in ocular optical coherence tomography

Thickness measurements derived from optical coherence tomography (OCT) images of the eye are a fundamental clinical and research metric, since they provide valuable information regarding the eye’s anatomical and physiological characteristics, and can assist in the diagnosis and monitoring of numerous ocular conditions. Despite the importance of these measurements, limited attention has been given to the methods used to estimate thickness in OCT images of the eye. Most current studies employing OCT use an axial thickness metric, but there is evidence that axial thickness measures may be biased by tilt and curvature of the image. In this paper, standard axial thickness calculations are compared with a variety of alternative metrics for estimating tissue thickness. These methods were tested on a data set of wide-field chorio-retinal OCT scans (field of view (FOV) 60° x 25°) to examine their performance across a wide region of interest and to demonstrate the potential effect of curvature of the posterior segment of the eye on the thickness estimates. Similarly, the effect of image tilt was systematically examined with the same range of proposed metrics. The results demonstrate that image tilt and curvature of the posterior segment can affect axial tissue thickness calculations, while alternative metrics, which are not biased by these effects, should be considered. This study demonstrates the need to consider alternative methods to calculate tissue thickness in order to avoid measurement error due to image tilt and curvature.OCIS codes: (100.0100) Image processing, (100.2960) Image analysis, (110.4500) Optical coherence tomography, (170.4470) Ophthalmology     

Robotic-arm-assisted flexible large field-of-view optical coherence tomography

Optical coherence tomography (OCT) is a three-dimensional non-invasive high-resolution imaging modality that has been widely used for applications ranging from medical diagnosis to industrial inspection. Common OCT systems are equipped with limited field-of-view (FOV) in both the axial depth direction (a few millimeters) and lateral direction (a few centimeters), prohibiting their applications for samples with large and irregular surface profiles. Image stitching techniques exist but are often limited to at most 3 degrees-of-freedom (DOF) scanning. In this work, we propose a robotic-arm-assisted OCT system with 7 DOF for flexible large FOV 3D imaging. The system consists of a depth camera, a robotic arm and a miniature OCT probe with an integrated RGB camera. The depth camera is used to get the spatial information of targeted sample at large scale while the RGB camera is used to obtain the exact position of target to align the image probe. Eventually, the real-time 3D OCT imaging is used to resolve the relative pose of the probe to the sample and as a feedback for imaging pose optimization when necessary. Flexible probe pose manipulation is enabled by the 7 DOF robotic arm. We demonstrate a prototype system and present experimental results with flexible tens of times enlarged FOV for plastic tube, phantom human finger, and letter stamps. It is expected that robotic-arm-assisted flexible large FOV OCT imaging will benefit a wide range of biomedical, industrial and other scientific applications.     

Reproducibility of optical coherence tomography airway imaging

Optical coherence tomography (OCT) is a promising imaging technique to evaluate small airway remodeling. However, the short-term insertion-reinsertion reproducibility of OCT for evaluating the same bronchial pathway has yet to be established. We evaluated 74 OCT data sets from 38 current or former smokers twice within a single imaging session. Although the overall insertion-reinsertion airway wall thickness (WT) measurement coefficient of variation (CV) was moderate at 12%, much of the variability between repeat imaging was attributed to the observer; CV for repeated measurements of the same airway (intra-observer CV) was 9%. Therefore, reproducibility may be improved by introduction of automated analysis approaches suggesting that OCT has potential to be an in-vivo method for evaluating airway remodeling in future longitudinal and intervention studies.OCIS codes: (170.0170) Medical optics and biotechnology, (170.4500) Optical coherence tomography     

MEMS scanning micromirror for optical coherence tomography

This paper describes an endoscopic-inspired imaging system employing a micro-electromechanical system (MEMS) micromirror scanner to achieve beam scanning for optical coherence tomography (OCT) imaging. Miniaturization of a scanning mirror using MEMS technology can allow a fully functional imaging probe to be contained in a package sufficiently small for utilization in a working channel of a standard gastroesophageal endoscope. This work employs advanced image processing techniques to enhance the images acquired using the MEMS scanner to correct non-idealities in mirror performance. The experimental results demonstrate the effectiveness of the proposed technique.OCIS codes: (230.4685) Optical microelectromechanical devices, (230.0230) Optical devices, (170.2150) Endoscopic imaging, (100.0100) Image processing