Polarization-sensitive Optical Coherence Tomography Research Articles

Glioma Cell Migration Dynamics in Brain Tissue Assessed by Multimodal Optical Imaging

Glioblastoma is a primary malignant brain tumor characterized by highly infiltrative glioma cells. Vasculature and white matter tracts are considered to be the preferred and fastest routes for glioma invasion through brain tissue. In this study, we systematically quantified the routes and motility of the U251 human glioblastoma cell line in mouse brain slices by multimodal imaging. Specifically, we used polarization-sensitive optical coherence tomography to delineate nerve fiber tracts while confocal fluorescence microscopy was used to image cell migration and brain vasculature. Somewhat surprisingly, we found that in mouse brain slices, U251 glioma cells do not follow white matter tracts but rather preferentially migrate along vasculature in both gray and white matter. In addition, U251 cell motility is ∼2-fold higher in gray matter than in white matter (91 vs. 43 μm2/h), with a substantial fraction (44%) of cells in both regions invading without close association with vasculature. Interestingly, within both regions, the rates of migration for the perivascular and televascular routes of invasion were indistinguishable. Furthermore, by imaging of local vasculature deformation dynamics during cell migration, we found that U251 cells are capable of exerting traction forces that locally pull on their environment, suggesting the applicability of a “motor-clutch”-based model for migration in vivo. Overall, by quantitatively analyzing the migration dynamics along the diverse pathways followed by invading U251 glioma cells as observed by our multimodal imaging approach, our studies suggest that effective antiinvasive strategies will need to simultaneously limit parallel routes of both perivascular and televascular invasion through both gray and white matter.

Hyperspectral optical coherence tomography for in vivo visualization of melanin in the retinal pigment epithelium.

Previous studies for melanin visualization in the retinal pigment epithelium (RPE) have exploited either its absorption properties (using photoacoustic tomography or photothermal optical coherence tomography [OCT]) or its depolarization properties (using polarization sensitive OCT). However, these methods are only suitable when the melanin concentration is sufficiently high. In this work, we present the concept of hyperspectral OCT for melanin visualization in the RPE when the concentration is low. Based on white light OCT, a hyperspectral stack of 27 wavelengths (440‐700 nm) was created in post‐processing for each depth‐resolved image. Owing to the size and shape of the melanin granules in the RPE, the variations in backscattering coefficient as a function of wavelength could be identified—a result which is to be expected from Mie theory. This effect was successfully identified both in eumelanin‐containing phantoms and in vivo in the low‐concentration Brown Norway rat RPE.

Evaluation of Retinal Pigment Epithelium Layer Change in Vogt-Koyanagi-Harada Disease With Multicontrast Optical Coherence Tomography.

Clinical evaluation of retinal pigment epithelium (RPE) change is important for the therapeutic management of chronic Vogt-Koyanagi-Harada (VKH) disease. We evaluated long-term change in the RPE layer in VKH disease, using near-infrared (NIR; 817 nm) images and autofluorescence images at 488 nm (short-wavelength [SW]-AF) and 785 nm (NIR-AF), and compared those images with images from multicontrast optical coherence tomography (MC-OCT). MC-OCT is capable of simultaneous measurement of OCT angiography, polarization-sensitive OCT, and standard OCT. We evaluated 24 eyes of 12 patients with chronic VKH disease. RPE changes were assessed using NIR, NIR-AF, SW-AF, and MC-OCT imaging performed from 6 to 48 months after disease onset. RPE-melanin-specific contrast OCT images were calculated using the dataset from MC-OCT. Granular hyper NIR-AF lesions were observed in 8 of 24 eyes (33%). Eyes with granular hyper NIR-AF lesions showed a sunset glow fundus appearance significantly more frequently than did eyes without such lesions (P < 0.0001). MC-OCT imaging confirmed that there was melanin accumulation at the RPE-Bruch's membrane band at the location of granular hyper NIR-AF lesions. Granular hyper NIR-AF lesions were unclear in SW-AF and color fundus images, but clearly detectable in NIR images. Areas of hyper NIR-AF lesions gradually decreased over time. Melanin accumulation in the RPE layer at the location of granular hyper NIR-AF lesions was confirmed with MC-OCT imaging. Long-term follow-up showed the reversible nature of this accumulation. MC-OCT is useful for the evaluation of change at the RPE layer in chronic VKH disease.

Optic axis uniformity as a metric to improve the contrast of birefringent structures and analyze the retinal nerve fiber layer in polarization-sensitive optical coherence tomography.

A new metric is used to improve the contrast of birefringent structures in biological tissue using polarization-sensitive optical coherence tomography. This metric, optic axis uniformity (OAxU), is based on the optic axis of birefringence and quantifies the uniformity of the optic axis direction. OAxU provides surprisingly strong contrast for fibrous structures such as muscle and the retinal nerve fiber layer (RNFL). We used OAxU for automatic segmentation of the RNFL in human eyes. From the segmentation, en face images of RNFL thickness and RNFL birefringence were created. The measured birefringence values are consistent with earlier reports.

Polarization-sensitive optical coherence tomography with a conical beam scan for the investigation of birefringence and collagen alignment in the human cervix.

By measuring the phase retardance of a cervical extracellular matrix, our in-house polarization-sensitive optical coherence tomography (PS-OCT) was shown to be capable of (1) mapping the distribution of collagen fibers in the non-gravid cervix, (2) accurately determining birefringence, and (3) measuring the distinctive depolarization of the cervical tissue. A conical beam scan strategy was also employed to explore the 3D orientation of the collagen fibers in the cervix by interrogating the samples with an incident light at 45° and successive azimuthal rotations of 0-360°. Our results confirmed previous observations by X-ray diffraction, suggesting that in the non-gravid human cervix collagen fibers adjacent to the endocervical canal and in the outermost areas tend to arrange in a longitudinal fashion whereas in the middle area they are oriented circumferentially. PS-OCT can assess the microstructure of the human cervical collagen in vitro and holds the potential to help us better understand cervical remodeling prior to birth pending the development of an in vivo probe.

Attenuation of stripe artifacts in optical coherence tomography images through wavelet-FFT filtering.

The use of polarization-maintaining (PM) fibers for polarization-sensitive optical coherence tomography (PS-OCT) can result in numerous image artifacts which degrade the reliability of birefringence measurements. Similar artifacts can also arise in conventional OCT, due to stray reflections from optical surfaces, a problem which is increasing in tandem with the steady rise in source coherence lengths. Here, a recently presented wavelet-FFT filter[Opt. Express 17(10), 8567 (2009).19434191] is combined with surface flattening displacement fields in order to suppress ghost artifacts following either a duplicate or inverse profile to that of the sample surface. In addition, horizontal coherence stripes originating from Fresnel reflections of optical components are suppressed in order to facilitate accurate surface detection. The result is an improved visualization of the phase-retardance profile within tissue, which may improve the reliability of curve-fitting methods for localized birefringence estimation. While the results are presented with a focus towards PS-OCT, the filtering method can also be applied to the removal of stray reflection artifacts in conventional OCT images.

Fiber-based polarization-sensitive optical coherence tomography of a minimalistic system configuration.

We present a very simple method of constructing a polarization-sensitive optical coherence tomography (PS-OCT) system. An ordinary fiber-based swept-source OCT system was reconfigured for PS-OCT by adding a long section of polarization-maintaining fiber in the sample arm. Two polarization modes of a large group-delay difference formed spatially distinguished polarization channels. The depth-encoded information on the polarization states was retrieved by an amplitude-based analysis. We found that our method provides an economic scheme of PS-OCT. It demonstrates that an ordinary OCT system can be easily reconfigured for PS-OCT imaging if it has sufficient margins in the imaging range.

Large Field of View Corneal Epithelium and Bowman's Layer Thickness Maps in Keratoconic and Healthy Eyes

To assess differences between epithelium thickness (ET) and Bowman's layer thickness (BLT) maps in keratoconic eyes and healthy eyes. Cross-sectional study. Setting: institutional. 47 patients (1 eye) with keratoconus (KC) and 20 healthy subjects (1 eye). epithelium and Bowman's layer measurements were performed by using custom-designed polarization-sensitive optical coherence tomography (PS-OCT) with a conical scanning optics design. En face corneal ET and BLT maps with a diameter of 11mm were computed. Main outcome measurements were mean ET and BLT of 25 sectors; the thinnest (minET, minBLT) and thickest sectors (maxET, maxBLT) were assessed. Ratios between thinnest/thickest sectors (R1) and between mean ET and BLT of the inferior temporal quadrant/superior nasal quadrant (R2) were calculated (R1ET, R1BLT; R2ET, R2BLT). Receiver operator characteristic (ROC) curve analysis was used to assess the diagnostic power of statistically different parameters. In healthy eyes, smooth ET maps were observed. KC eyes showed a "doughnut pattern." The BLT maps of healthy eyes had a smooth appearance, but highly irregular "moth"-like damage pattern could be observed in keratoconic eyes. Highest area under the curve values were found for the thinnest sector of the BLT map, the R1ET, and the thinnest sector of the ET map. PS-OCT imaging enables the visualization of significant differences of the corneal epithelium and the Bowman's layer in en face maps covering almost the entire cornea. ET and BLT profiles could clearly show their diagnostic importance for the distinguishing of keratoconic eyes and healthy eyes.

In vivo multifunctional optical coherence tomography at the periphery of the lungs.

Remodeling of tissue, such as airway smooth muscle (ASM) and extracellular matrix, is considered a key feature of airways disease. No clinically accepted diagnostic method is currently available to assess airway remodeling or the effect of treatment modalities such as bronchial thermoplasty in asthma, other than invasive airway biopsies. Optical coherence tomography (OCT) generates cross-sectional, near-histological images of airway segments and enables identification and quantification of airway wall layers based on light scattering properties only. In this study, we used a custom motorized OCT probe that combines standard and polarization sensitive OCT (PS-OCT) to visualize birefringent tissue in vivo in the airway wall of a patient with severe asthma in a minimally invasive manner. We used optic axis uniformity (OAxU) to highlight the presence of uniformly arranged fiber-like tissue, helping visualizing the abundance of ASM and connective tissue structures. Attenuation coefficient images of the airways are presented for the first time, showing superior architectural contrast compared to standard OCT images. A novel segmentation algorithm was developed to detect the surface of the endoscope sheath and the surface of the tissue. PS-OCT is an innovative imaging technique that holds promise to assess airway remodeling including ASM and connective tissue in a minimally invasive, real-time manner.

Comparison of Intensity- and Polarization-based Contrast in Amyloid-beta Plaques as Observed by Optical Coherence Tomography

One key hallmark of Alzheimer’s disease (AD) is the accumulation of extracellular amyloid-beta protein in cortical regions of the brain. For a definitive diagnosis of AD, post-mortem histological analysis, including sectioning and staining of different brain regions, is required. Here, we present optical coherence tomography (OCT) as a tissue-preserving imaging modality for the visualization of amyloid-beta plaques and compare their contrast in intensity- and polarization-sensitive (PS) OCT. Human brain samples of eleven patients diagnosed with AD were imaged. Three-dimensional PS-OCT datasets were acquired and plaques were manually segmented in 500 intensity and retardation cross-sections per patient using the freely available ITK-SNAP software. The image contrast of plaques was quantified. Histological staining of tissue sections from the same specimens was performed to compare OCT findings against the gold standard. Furthermore, the distribution of plaques was evaluated for intensity-based OCT, PS-OCT and the corresponding histological amyloid-beta staining. Only 5% of plaques were visible in both intensity and retardation segmentations, suggesting that different types of plaques may be visualized by the two OCT contrast channels. Our results indicate that multicontrast OCT imaging might be a promising approach for a tissue-preserving visualization of amyloid-beta plaques in AD.

Parallel detection of Jones-matrix elements in polarization-sensitive optical coherence tomography.

The polarization properties of a sample can be characterized using a Jones matrix. To measure the Jones matrix without assumptions of the sample, two different incident states of polarization are usually used. This requirement often causes certain drawbacks in polarization-sensitive optical coherence tomography (PS-OCT), e.g., a decrease in the effective A-scan rate or axial depth range, if a multiplexing scheme is used. Because both the A-scan rate and axial depth range are important for clinical applications, including the imaging of an anterior eye segment, a new PS-OCT method that does not have these drawbacks is desired. Here, we present a parallel-detection approach that maintains the same A-scan rate and axial measurement range as conventional OCT. The interferometer consists of fiber-optic components, most of which are polarization-maintaining components with fast-axis blocking free from polarization management. When a parallel detection is implemented using swept-source OCT (SS-OCT), synchronization between the A-scans and synchronization between the detection channels have critical effects on the Jones-matrix measurement. Because it is difficult to achieve perfect synchronization using only hardware, we developed a solution using a numerical correction with signals from a static mirror. Using the developed system, we demonstrate the imaging of an anterior eye segment from the cornea to the back surface of the crystalline lens.

Catheter-Based Polarization Sensitive Optical Coherence Tomography Using Similar Mueller Matrix Method.

Research of catheter-based polarization sensitive optical coherence tomography (PS-OCT) is a challenging field. In this paper, we present a new polarization determination method, similar Mueller matrix (SMM) method, for a catheter-based PS-OCT system using a standard clinical catheter probe with an outer diameter of 0.9 mm. The SMM method can remove the diattenuation and depolarization compositions by polar decomposition. By constructing the similarity between the measured Mueller matrices and sample matrices, the phase retardance of the sample can be determined from the trace of the measured matrices. In the experiments, we find that images processed by the SMM method without any averaging or phase correction have a better polarization contrast of multiple biological tissues than those by the Jones matrix based method. We also preliminarily achieve phase retardance imaging of the ex vivo porcine cardiac blood vessel. Compared with the Jones matrix based method, the presented SMM method can provide a more robust birefringence imaging of biological tissues under low signal-to-noise ratio, depolarization, diattenuation, and phase instability. The SMM method has a potential to become a widely accepted polarization determination method for catheter-based PS-OCT.

Depth-resolved birefringence imaging of collagen fiber organization in the human oral mucosa in vivo.

Stromal collagen organization has been identified as a potential prognostic indicator in a variety of cancers and other diseases accompanied by fibrosis. Changes in the connective tissue are increasingly considered for grading dysplasia and progress of oral squamous cell carcinoma, investigated mainly ex vivo by histopathology. In this study, polarization-sensitive optical coherence tomography (PS-OCT) with local phase retardation imaging is used for the first time to visualize depth-resolved (i.e., local) birefringence of healthy human oral mucosa in vivo. Depth-resolved birefringence is shown to reveal the expected local collagen organization. To demonstrate proof-of-principle, 3D image stacks were acquired at labial and lingual locations of the oral mucosa, chosen as those most commonly affected by cancerous alterations. To enable an intuitive evaluation of the birefringence images suitable for clinical application, color depth-encoded en-face projections were generated. Compared to en-face views of intensity or conventional cumulative phase retardation, we show that this novel approach offers improved visualization of the mucosal connective tissue layer in general, and reveals the collagen fiber architecture in particular. This study provides the basis for future prospective pathological and comparative in vivo studies non-invasively assessing stromal changes in conspicuous and cancerous oral lesions at different stages.

Slope-based segmentation of articular cartilage using polarization-sensitive optical coherence tomography phase retardation image.

.A segmentation method based on phase retardation measurements from polarization-sensitive optical coherence tomography (PS-OCT) is developed to differentiate the structural zones of articular cartilage. The organization of collagen matrix in articular cartilage varies over the different structural zones, generating different tissue birefringence. Analyzing the slope of the accumulated phase retardation at different depths can detect the variation in tissue birefringence and be used to segment the structural zones. The method is validated on phantoms composed of layers of different materials. Articular cartilage samples from adult swine are segmented with the method. The characteristics in each segmented zone are also examined by histology and high-resolution second-harmonic generation imaging, showing distinctive properties that match with the anatomical structure of articular cartilage. The segmentation algorithm is also applied on PS-OCT images acquired at multiple illumination angles, where the angular dependence of tissue birefringence in the deep zone is detected. This method offers a noninvasive imaging approach to differentiating the structural zones of articular cartilage, as well as a quantification approach based on the phase retardation measurements of PS-OCT. This method has great potential in studying depth-related progression of cartilage degeneration.

IMAGING OF VITELLIFORM MACULAR LESIONS USING POLARIZATION-SENSITIVE OPTICAL COHERENCE TOMOGRAPHY.

To examine the involvement of the retinal pigment epithelium (RPE) in the presence of vitelliform macular lesions (VML) in Best vitelliform macular dystrophy (BVMD), autosomal recessive bestrophinopathy, and adult-onset vitelliform macular degeneration using polarization-sensitive optical coherence tomography (PS-OCT). A total of 35 eyes of 18 patients were imaged using a PS-OCT system and blue light fundus autofluorescence imaging. Pathogenic mutations in the BEST1 gene, 3 of which were new, were detected in all patients with BVMD and autosomal recessive bestrophinopathy. Polarization-sensitive optical coherence tomography showed a characteristic pattern in all three diseases with nondepolarizing material in the subretinal space consistent with the yellowish VML seen on funduscopy with a visible RPE line below it. A focal RPE thickening was seen in 26 eyes under or at the edge of the VML. Retinal pigment epithelium thickness outside the VML was normal or mildly thinned in patients with BVMD and adult-onset vitelliform macular degeneration but was diffusely thinned or atrophic in patients with autosomal recessive bestrophinopathy. Patients with autosomal recessive bestrophinopathy showed sub-RPE fibrosis alongside the subretinal VML. Polarization-sensitive optical coherence tomography was more reliable in assessing the localization and the integrity of the RPE than spectral domain OCT alone. On spectral domain OCT, identification of the RPE was not possible in 19.4% of eyes. Polarization-sensitive optical coherence tomography allowed for definite identification of the location of VML in respect to the RPE in all eyes, since it provides a tissue-specific contrast. Polarization-sensitive optical coherence tomography confirms in vivo the subretinal location of VML and is useful in the assessment of RPE integrity.

Evaluation of focal damage in the retinal pigment epithelium layer in serous retinal pigment epithelium detachment

The purpose of this study was to evaluate focal damage in the retinal pigment epithelium (RPE) layer in serous retinal pigment epithelium detachment (PED) with multi-contrast optical coherence tomography (OCT), which is capable of simultaneous measurement of OCT angiography, polarization-sensitive OCT and standard OCT images. We evaluated 37 eyes with age-related macular degeneration that had serous PED. Focal RPE damage was indicated by hyper-transmission beneath the RPE-Bruch’s membrane band in standard OCT images. Distribution of RPE melanin was calculated using the dataset from multi-contrast OCT. Twenty-four points with hyper-transmission were detected in 21 of the 37 eyes. Standard OCT images failed to show disruption of the RPE-Bruch’s membrane band at 5 of the 24 hyper-transmission points. Conversely, multi-contrast OCT images clearly showed melanin defects in the RPE-Bruch’s membrane band at all points. Areas of melanin defects with disruption of the RPE-Bruch’s membrane band were significantly larger than those without disruption. The volume of intraretinal hyper-reflective foci was significantly larger in eyes with hyper-transmission than that in eyes without hyper-transmission. Multi-contrast OCT is more sensitive than standard OCT for displaying changes at the RPE-Bruch’s membrane band when there are small areas of RPE damage.

Single input state polarization-sensitive optical coherence tomography with high resolution and polarization distortion correction.

In single input state polarization-sensitive optical coherence tomography (PS-OCT) with high resolution, the imperfections of quarter-wave plate (QWP) and the sensitivity roll-off mismatch between the two detection channels cause unpredictable polarization distortion. We present a correction method based on the Jones matrix modeling of the system. In a single input PS-OCT system working at 840 nm with an axial resolution of ~2.3 μm, the method yielded better estimation of retardation and optic axis orientation with significantly reduced noise level, especially in weakly birefringent samples. Numerical simulations and quantitative imaging of a sample of known birefringence were performed to validate the performance. We further demonstrate the advantages of our approach with birefringence imaging of swine retina, rat aortic wall, and rat esophageal mucosa for potential clinical applications.

Identification of Human Pathological Mitral Chordae Tendineae Using Polarization-sensitive Optical Coherence Tomography

Defects of the mitral valve complex imply heart malfunction. The chordae tendineae (CTs) are tendinous strands connecting the mitral and tricuspid valve leaflets to the papillary muscles. These CTs are composed of organized, wavy collagen bundles, making them a strongly birefringent material. Disorder of the collagen structure due to different diseases (rheumatic, degenerative) implies the loss or reduction of tissue birefringence able to be characterized with Polarization Sensitive Optical Coherence Tomography (PS-OCT). PS-OCT is used to discriminate healthy from diseased chords, as the latter must be excised and replaced in clinical conventional interventions. PS-OCT allows to quantify birefringence reduction in human CTs affected by degenerative and rheumatic pathologies. This tissue optical property is proposed as a diagnostic marker for the identification of degradation of tendinous chords to guide intraoperative mitral valve surgery.

Identification and Quantification of the Angiofibrotic Switch in Neovascular AMD.

We quantify volumetric changes of subretinal hyperreflective material (SHRM) and determine the conversion toward subretinal fibrosis, the angiofibrotic switch, under anti-VEGF therapy using polarization-sensitive optical coherence tomography (PS-OCT). A total of 50 eyes of 50 patients with treatment-naïve neovascular age-related macular degeneration (AMD) were included in this prospective observational study: 26 diagnosed with type 1 choroidal neovascularization (CNV), seven with type 2 CNV, 11 with mixed type CNV, three with a retinal angiomatous proliferation (RAP) lesion and three with a polypoidal choroidal vasculopathy (PCV). Patients were imaged at baseline and at the end of the loading phase (after treatment with three intravitreal anti-VEGF injections) using a PS-OCT system with a scanning angle of 30° × 30° and a scan pattern of 1024 × 250 A-scans. The device is capable of detecting fibrosis based on birefringence and the RPE based on depolarization. The volume of SHRM was quantified by manual delineation in each PS-OCT B-scan and interpolation between B-scans using proprietary reading center certified software. The occurrence of fibrosis detected by PS-OCT was compared to the clinical presentation of subretinal fibrosis. Of 50 eyes, 28 had SHRM at baseline. Seven of these eyes had subretinal fibrosis within 3 months, six of which could be detected unambiguously based on PS-OCT imaging. SHRM thickness and volume at month 3 (P = 0.001 and P = 0.02) were significantly larger and the reduction of SHRM thickness and volume (P = 0.002 and P = 0.027) in response to therapy were significantly less pronounced in eyes with fibrosis. SHRM volume decreases significantly under anti-VEGF therapy. However, lesions unresponsive to therapy may progress to fibrosis as early as 3 months. Reduction in SHRM thickness may be a prognostic marker for treatment response.

Visual Function and Integrity of Depolarization Measured by Polarization-sensitive Optical Coherence Tomography in Patients With Neovascular Age-related Macular Degeneration

Visual Function and Integrity of Depolarization Measured by Polarization-sensitive Optical Coherence Tomography in Patients With Neovascular Age-related Macular Degeneration