Retinal nerve fiber layer birefringence evaluated with polarization sensitive spectral domain OCT and scanning laser polarimetry: A comparison

A polarization‐sensitive spectral domain optical coherence tomography (PS‐SD‐OCT) system is used to measure phase retardation and birefringence of the human retinal nerve fiber layer (RNFL) in vivo. The instrument records three parameters simultaneously: intensity, phase retardation and optic‐axis orientation. 3D data sets are recorded in the optic nerve‐head area of a healthy and a glaucomatous eye, and the results are presented in various ways: En‐face phase‐retardation maps of the RNFL are generated from the recorded 3D data and results are compared with scanning laser polarimetry (SLP). The depth information provided by OCT is used to segment the RNFL in the intensity image and measure the RNFL thickness. From the retardation and thickness data, 2D birefringence maps of the RNFL are derived. Circumpapillary plots of RNFL retardation and thickness obtained by PS‐SD‐OCT are quantitatively compared with those obtained by SLP. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Measuring polarization changes in the human outer retina with polarization‐sensitive optical coherence tomography

Morphological changes in the outer retina such as drusen are established biomarkers to diagnose age‐related macular degeneration. However, earlier diagnosis might be possible by taking advantage of more subtle changes that accompany tissues that bear polarization‐altering properties. To test this hypothesis, we developed a method based on polarization‐sensitive optical coherence tomography with which volumetric data sets of the macula were obtained from 10 young (<25 years) and 10 older (>54 years) subjects. All young subjects and 5 of the older subjects had retardance values induced by the retinal pigment epithelium and Bruch's membrane (RPE‐BM) complex that were just above the noise floor measurement (5°‐13° at 840 nm). In contrast, elevated retardance, up to 180°, was observed in the other 5 older subjects. Analysis of the degree of polarization uniformity (DOPU) demonstrates that reduced DOPU (<0.4) in the RPE is associated with elevated double pass phase retardation (DPPR) below the RPE‐BM complex, suggesting that the observed elevated DPPR in older subjects is the result of increased scattering or polarization scrambling. Collectively, our measurements show that the outer retina can undergo dramatic change in its polarization properties with age, and in some cases still retain its clinically normal appearance.      

A feasibility study of OCT for anatomical and vascular phenotyping of mouse embryo

The embryo phenotyping of genetic murine model is invaluable when investigating functions of genes underlying embryonic development and birth defect. Although traditional imaging technologies such as ultrasound are very useful for evaluating phenotype of murine embryos, the use of advanced techniques for phenotyping is desirable to obtain more information from genetic research. This letter tests the feasibility of optical coherence tomography (OCT) as a high‐throughput phenotyping tool for murine embryos. Three‐dimensional OCT imaging is performed for live and cleared mouse embryos in the late developmental stage (embryonic day 17.5). By using a dynamic focusing method and OCT angiography (OCTA) approach, our OCT imaging of the embryo exhibits rapid and clean visualization of organ structures deeper than 5 mm and complex microvasculature of perfused blood vessels in the murine embryonic body. This demonstration suggests that OCT imaging can be useful for comprehensively assessing embryo anatomy and angiography of genetically engineered mice.     

Multicontrast endomyocardial imaging by single‐channel high‐resolution cross‐polarization optical coherence tomography

A single‐channel high‐resolution cross‐polarization (CP) optical coherence tomography (OCT) system is presented for multicontrast imaging of human myocardium in one‐shot measurement. The intensity and functional contrasts, including the ratio between the cross‐ and co‐polarization channels as well as the cumulative retardation, are reconstructed from the CP‐OCT readout. By comparing the CP‐OCT results with histological analysis, it is shown that the system can successfully delineate microstructures in the myocardium and differentiate the fibrotic myocardium from normal or ablated myocardium based on the functional contrasts provided by the CP‐OCT system. The feasibility of using A‐line profiles from the 2 orthogonal polarization channels to identify fibrotic myocardium, normal myocardium and ablated lesion is also discussed.      

Quantitative depolarization measurements for fiber‐based polarization‐sensitive optical frequency domain imaging of the retinal pigment epithelium

A full quantitative evaluation of the depolarization of light may serve to assess concentrations of depolarizing particles in the retinal pigment epithelium and to investigate their role in retinal diseases in the human eye. Optical coherence tomography and optical frequency domain imaging use spatial incoherent averaging to compute depolarization. Depolarization depends on accurate measurements of the polarization states at the receiver but also on the polarization state incident upon and within the tissue. Neglecting this dependence can result in artifacts and renders depolarization measurements vulnerable to birefringence in the system and in the sample. In this work, we discuss the challenges associated with using a single input polarization state and traditional depolarization metrics such as the degree‐of‐polarization and depolarization power. We demonstrate quantitative depolarization measurements based on Jones vector synthesis and polar decomposition using fiber‐based polarization‐sensitive optical frequency domain imaging of the retinal pigment epithelium in a human eye.      

Use of 2D images of depth and integrated reflectivity to represent the severity of demineralization in cross‐polarization optical coherence tomography

Several studies have demonstrated the potential of cross‐polarization optical coherence tomography (CP‐OCT) to quantify the severity of early caries lesions (tooth decay) on tooth surfaces. The purpose of this study is to show that 2D images of the lesion depth and the integrated reflectivity can be used to accurately represent the severity of early lesions. Simulated early lesions of varying severity were produced on tooth samples using simulated lesion models. Methods were developed to convert the 3D CP‐OCT images of the samples to 2D images of the lesion depth and lesion integrated reflectivity. Calculated lesion depths from OCT were compared with lesion depths measured from histological sections examined using polarized light microscopy. The 2D images of the lesion depth and integrated reflectivity are well suited for visualization of early demineralization. (© 2013 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)     

Pixel classification method in optical coherence tomography for tumor segmentation and its complementary usage with OCT microangiography

A novel machine‐learning method to distinguish between tumor and normal tissue in optical coherence tomography (OCT) has been developed. Pre‐clinical murine ear model implanted with mouse colon carcinoma CT‐26 was used. Structural‐image‐based feature sets were defined for each pixel and machine learning classifiers were trained using “ground truth” OCT images manually segmented by comparison with histology. The accuracy of the OCT tumor segmentation method was then quantified by comparing with fluorescence imaging of tumors expressing genetically encoded fluorescent protein KillerRed that clearly delineates tumor borders. Because the resultant 3D tumor/normal structural maps are inherently co‐registered with OCT derived maps of tissue microvasculature, the latter can be color coded as belonging to either tumor or normal tissue. Applications to radiomics‐based multimodal OCT analysis are envisioned.      

Development of a polarization imaging method to detect paraffin-embedded pathology tissues before applying other techniques

The present article describes the development of a technique, applied to paraffin-embedded tissues, which uses three different wavelengths of monochromatic light (λ1 = 445 nm, λ2 = 540 nm and λ3 = 660 nm) for the measures of the degree of polarization, degree of linear polarization, degree of circular polarization and birefringence, all obtained from measurements of Stokes parameters by using polarized light. The goal of this study was to detect changes in developing embryonic mouse eye when pregnant mice fed diets without folic acid for variable periods compared with a healthy control group. We present a biomedical diagnostic technique based on polarized light detection applied to paraffin-embedded tissues to visualize the structural damage to aid us in the diagnosis before applying other techniques. Through this method, we can visualize and identify which parts of the tissue were altered with respect to the control group.     

Real-time measurement of repetitive micro bulk motion vector and motion noise removal in optical coherence tomography angiography

In this work, we developed a motion estimation and correction method which real-time obtained the direction and displacement of repetitive micro bulk motion (such as cardiac and respiratory motion) on an SS-OCT system without additional tracking hardware, and reduced the motion noise in optical coherence tomography angiography (OCTA). In the approach, the direction of repetitive micro bulk motion was considered fixed, and proportional relationships between the motion components in three directions were determined; Then we performed one-dimension cross-correlation to obtain depth displacement which was further used to obtain other two motion components, and greatly reduced the computation; The processing speed on a graphic processing unit was 478 pairs of B-Scans per second, and the measurement range was larger than the range of the angiogram-based methods. Lastly, corrupt angiograms were recovered by adaptive scan protocol, and reduced acquisition time in comparison with the previous work.     

Comparative study of deep neural networks with unsupervised Noise2Noise strategy for noise reduction of optical coherence tomography images

As a powerful diagnostic tool, optical coherence tomography (OCT) has been widely used in various clinical setting. However, OCT images are susceptible to inherent speckle noise that may contaminate subtle structure information, due to low-coherence interferometric imaging procedure. Many supervised learning-based models have achieved impressive performance in reducing speckle noise of OCT images trained with a large number of noisy-clean paired OCT images, which are not commonly feasible in clinical practice. In this article, we conducted a comparative study to investigate the denoising performance of OCT images over different deep neural networks through an unsupervised Noise2Noise (N2N) strategy, which only trained with noisy OCT samples. Four representative network architectures including U-shaped model, multi-information stream model, straight-information stream model and GAN-based model were investigated on an OCT image dataset acquired from healthy human eyes. The results demonstrated all four unsupervised N2N models offered denoised OCT images with a performance comparable with that of supervised learning models, illustrating the effectiveness of unsupervised N2N models in denoising OCT images. Furthermore, U-shaped models and GAN-based models using UNet network as generator are two preferred and suitable architectures for reducing speckle noise of OCT images and preserving fine structure information of retinal layers under unsupervised N2N circumstances.     

An automated framework to quantify areas of regional ischemia in retinal vascular diseases with OCT angiography

In this observational and cross‐sectional study, capillary nonperfusion (CNP) and vascular changes in branch retinal vein occlusion (BRVO, sample size [n] = 26) and choroidal neovascularization (CNV, n = 29) were evaluated. Subjects underwent imaging using Optical coherence tomography angiography (Angiovue OCTA, RTVue XR, Optovue Inc., Fremont, California). Local fractal analysis was applied to the OCTA images of superficial, deep and choriocapillaris layer. CNP area (BRVO eyes) and vascular parameters were computed using local fractal‐based method. Sensitivity and specificity of vascular parameters were assessed with receiver operating characteristics curve. Automated CNP area showed excellent agreement with manually quantified CNP areas in both superficial (intraclass coefficient [ICC] = 0.96) and deep (ICC = 0.96) layers. BRVO eyes showed significantly altered (P < .05) vascular parameters in both superficial and deep layer as compared to normal eyes (n = 30). CNVM eyes had significantly higher capillary free zones (P < .001) as compared to normal eyes. In normal vs BRVO eyes, vessel density and spacing between the large vessels had similar area under the curve (AUC) (P > .05) in both superficial (0.97 and 0.97, respectively) and deep layer (0.99 and 0.98, respectively). Further, capillary free zones showed high AUC (0.92) in differentiating CNV eyes from normal eyes.      

Three-dimensional spatial reconstruction of coronary arteries based on fusion of intravascular optical coherence tomography and coronary angiography

We present a three-dimensional (3D) spatial reconstruction of coronary arteries based on fusion of intravascular optical coherence tomography (IVOCT) and digital subtraction angiography (DSA). Centerline of vessel in DSA images is exacted by multi-scale filtering, adaptive segmentation, morphology thinning and Dijkstra's shortest path algorithm. We apply the cross-correction between lumen shapes of IVOCT and DSA images and match their stenosis positions to realize co-registration. By matching the location and tangent direction of the vessel centerline of DSA images and segmented lumen coordinates of IVOCT along pullback path, 3D spatial models of vessel lumen are reconstructed. Using 1121 distinct positions selected from eight vessels, the correlation coefficient between 3D IVOCT model and DSA image in measuring lumen radius is 0.94% and 97.7% of the positions fall within the limit of agreement by Bland–Altman analysis, which means that the 3D spatial reconstruction IVOCT models and DSA images have high matching level.     

Investigation of the permeability and optical clearing ability of different analytes in human normal and cancerous breast tissues by spectral domain OCT

The potential of OCT applied for early breast cancer detection attracted significant efforts. The permeability coefficients and the percentages of OCT signal enhancement for normal and cancerous breast tissues have been investigated from the experiments of 20% glucose, 40% glucose, and 20% mannitol in vitro. Obtained results indicate that the permeability coefficient in breast cancer tissue is prominently larger than that in normal breast tissue, while the optical clearing effect is just the reverse to that for each agent. The results suggest that OCT has the ability to distinguish cancer tissue from different aspect. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

‘Go with the flow ’: A review of methods and advancements in blood flow imaging

Physics has delivered extraordinary developments in almost every facet of modern life. From the humble thermometer and stethoscope to X‐Ray, CT, MRI, ultrasound, PET and radiotherapy, our health has been transformed by these advances yielding both morphological and functional metrics. Recently high resolution label‐free imaging of the microcirculation at clinically relevant depths has become available in the research domain. In this paper, we present a comprehensive review on current imaging techniques, state‐of‐the‐art advancements and applications, and general perspectives on the prospects for these modalities in the clinical realm. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Retinal imaging with optical coherence tomography and low‐loss adaptive optics using a 2.8‐mm beam size

As data acquisition for retinal imaging with optical coherence tomography (OCT) becomes faster, efficient collection of photons becomes more important to maintain image quality. One approach is to use a larger aperture at the eye's pupil to collect more photons that have been reflected from the retina. A 2.8‐mm beam diameter system with only seven reflecting surfaces was developed for low‐loss retinal imaging. The larger beam size requires defocus and astigmatism correction, which was done in a closed loop adaptive optics method using a Shack‐Hartmann wavefront sensor and a deformable mirror (DM) with 140 actuators and a ±2.75 μm stroke. This DM facilitates defocus correction ranging from approximately ?3 D to +3 D. Comparing the new system with a standard 1.2‐mm system on a model eye, a signal‐to‐noise gain of 4.5 dB and a 2.3 times smaller speckle size were measured. Measurements on the retinas of five subjects showed even better results, with increases in dynamic range up to 13 dB. Note that the new sample arm only occupies 30 cm × 60 cm, which makes it highly suitable for imaging in a clinical environment. Figure: B‐scan images obtained over a width of 8 deg from the right eye of a 31‐year‐old Caucasian male. While the left side was imaged with a standard 1.2‐mm OCT system, the right side was imaged with the 2.8‐mm system. Both images were collected with the same integration time and incident power, after correction of aberrations. Using the dynamic range within the images, which is determined by comparing the highest pixel value to the noise floor, a difference in dynamic range of 10.8 dB was measured between the two systems.      

Spectral correction for handheld optoacoustic imaging by means of near‐infrared optical tomography in reflection mode

In vivo imaging of tissue/vasculature oxygen saturation levels is of prime interest in many clinical applications. To this end, the feasibility of combining two distinct and complementary imaging modalities is investigated: optoacoustics (OA) and near‐infrared optical tomography (NIROT), both operating noninvasively in reflection mode. Experiments were conducted on two optically heterogeneous phantoms mimicking tissue before and after the occurrence of a perturbation. OA imaging was used to resolve submillimetric vessel‐like optical absorbers at depths up to 25 mm, but with a spectral distortion in the OA signals. NIROT measurements were utilized to image perturbations in the background and to estimate the light fluence inside the phantoms at the wavelength pair (760 nm, 830 nm). This enabled the spectral correction of the vessel‐like absorbers' OA signals: the error in the ratio of the absorption coefficient at 830 nm to that at 760 nm was reduced from 60%‐150% to 10%‐20%. The results suggest that oxygen saturation (SO ₂) levels in arteries can be determined with <10% error and furthermore, that relative changes in vessels' SO ₂ can be monitored with even better accuracy. The outcome relies on a proper identification of the OA signals emanating from the studied vessels.      

Indocyanine green-enhanced multimodal photoacoustic microscopy and optical coherence tomography molecular imaging of choroidal neovascularization

Photoacoustic microscopy (PAM) has great potential for visualization of the microvasculature with high spatial resolution and contrast. Early detection and differentiation of newly developed blood vessels named choroidal neovascularization (CNV) from normal vasculature remains a challenge in ophthalmology. Exogenous contrast agents can assist with improving PAM sensitivity, leading to differentiation of CNV. Here, an FDA-approved indocyanine green (ICG) was utilized as a PAM contrast agent. ICG was conjugated with RGD peptides, allowing the ICG to bind to the integrin expressed in CNV. Molecular PAM imaging showed that ICG-RGD can target CNV for up to 5 days post intravenous administration in living rabbits with a model of CNV. The PAM image sensitivity and image contrast were significantly enhanced by 15-fold at 24 h post-injection. Overall, the presented approach demonstrates the possibility of targeted ICG to be employed in PAM molecular imaging, allowing more precise evaluation of neovascularization.     

Activity assessment of root caries lesions with thermal and near‐IR imaging methods

The purpose of this study was to evaluate thermal and near‐infrared (NIR) reflectance imaging methods for the assessment of the activity of root caries lesions. In addition, changes in the lesion structure were monitored with polarization sensitive optical coherence tomography (PS‐OCT). Artificial bovine and natural root caries lesions were imaged with PS‐OCT, and their dehydration rate was measured with thermal and NIR cameras. The lesion activity of the natural root caries samples was also assessed by two clinicians by conventional means according to ICDAS II guidelines. The thickness of the highly mineralized transparent surface layer measured using PS‐OCT increased and the area enclosed by the time‐temperature curve, ΔQ, measured with thermal imaging decreased significantly with longer periods of remineralization in simulated dentin lesions, but the NIR reflectance intensity differences, ΔI, failed to show any significant relationship with the degree of remineralization. The PS‐OCT algorithm for the automated assessment of remineralization successfully detected the highly mineralized surface layer on both natural and simulated lesions. Thermal imaging provided the most accurate diagnosis of root caries lesion activity. These results demonstrate that thermal imaging and PS‐OCT may be ideally suited for the nondestructive root caries lesion activity during a clinical examination.