Nondestructive label-free detection of peritumoral white matter damage using cross-polarization optical coherence tomography.

Introduction: To improve the quality of brain tumor resections, it is important to differentiate zones with myelinated fibers destruction from tumor tissue and normal white matter. Optical coherence tomography (OCT) is a promising tool for brain tissue visualization and in the present study, we demonstrate the ability of cross-polarization (CP) OCT to detect damaged white matter and differentiate it from normal and tumor tissues. Materials and methods: The study was performed on 215 samples of brain tissue obtained from 57 patients with brain tumors. The analysis of the obtained OCT data included three stages: 1) visual analysis of structural OCT images; 2) quantitative assessment based on attenuation coefficients estimation in co- and cross-polarizations; 3) building of color-coded maps with subsequent visual analysis. The defining characteristics of structural CP OCT images and color-coded maps were determined for each studied tissue type, and then two classification tests were passed by 8 blinded respondents after a training. Results: Visual assessment of structural CP OCT images allows detecting white matter areas with damaged myelinated fibers and differentiate them from normal white matter and tumor tissue. Attenuation coefficients also allow distinguishing all studied brain tissue types, while it was found that damage to myelinated fibers leads to a statistically significant decrease in the values of attenuation coefficients compared to normal white matter. Nevertheless, the use of color-coded optical maps looks more promising as it combines the objectivity of optical coefficient and clarity of the visual assessment, which leads to the increase of the diagnostic accuracy of the method compared to visual analysis of structural OCT images. Conclusions: Alteration of myelinated fibers causes changes in the scattering properties of the white matter, which gets reflected in the nature of the received CP OCT signal. Visual assessment of structural CP OCT images and color-coded maps allows differentiating studied tissue types from each other, while usage of color-coded maps demonstrates higher diagnostic accuracy values in comparison with structural images (F-score = 0.85-0.86 and 0.81, respectively). Thus, the results of the study confirm the potential of using OCT as a neuronavigation tool during resections of brain tumors.

Brain white matter morphological structure correlation with its optical properties estimated from optical coherence tomography (OCT) data.

A pilot post-mortem study identifies a strong correlation between the attenuation coefficient estimated from the OCT data and some morphological features of the sample, namely the number of nuclei in the field of view of the histological image and the fiber structural parameter introduced in the study to quantify the difference in the myelinated fibers arrangements. The morphological features were identified from the histopathological images of the sample taken from the same locations as the OCT images and stained with the immunohistochemical (IHC) staining specific to the myelin. It was shown that the linear regression of the IHC quantitative characteristics allows adequate prediction of the attenuation coefficient of the sample. This discovery opens the opportunity for the usage of the OCT as a neuronavigation tool.

Lymph vessels visualization from optical coherence tomography data using depth-resolved attenuation coefficient calculation.

Multimodal optical coherent tomography grows popularity with researchers and clinicians over the past decade. One of the modalities is lymphangiography, which allows visualization of the lymphatic vessel networks within optical coherence tomography (OCT) imaging volume. In the present study, it is shown that lymphatic vessel visualization obtained from the depth-resolved attenuation coefficient distributions, corrected for the noise, shows improved contrast and detail in comparison with previously proposed approaches. We also argue that the two most popular approaches for lymphatic vessel visualization, namely simple intensity thresholding and vesselness calculation based on local Hessian matrix eigenvalues, imply different definitions of the lymphatic vessel's appearance in the OCT volume and lead to the different networks.

Optical coefficients as tools for increasing the optical coherence tomography contrast for normal brain visualization and glioblastoma detection.

The methods used for digital processing of optical coherence tomography (OCT) and crosspolarization (CP) OCT images are focused on improving the contrast ratio of native structural OCT images. Such advances are particularly important for the intraoperative detection of glioma margins where the visual assessment of OCT images can be difficult and lead to errors. The aim of the study was to investigate the application of optical coefficients obtained from CP OCT data for the differentiation of glial tumorous tissue from a normal brain. Pseudocolor en-face OCT maps based on two optical coefficients (the commonly used rate of attenuation in the cochannel, and in addition, the interchannel attenuation difference) were constructed for normal rat brain coronal cross sections and for brains with a 101.8 rat glioblastoma model. It was shown that the use of optical coefficients significantly increased the available information from the OCT data in comparison with unprocessed images. As a result, this allowed contrasting of the white matter from the gray matter and tumorous tissue ex vivo, and for this purpose, the interchannel attenuation difference worked better. The interchannel attenuation difference values of white matter were at least seven and two times higher than corresponding values of the cortex and tumorous tissue, whereas the same parameter for cochannel attenuation coefficient values of white matter are about 4 and 1.4. However, quantitative analysis shows that both coefficients are suitable for the purpose of glioblastoma detection from normal brain tissue regardless of whether a necrotic component was present (in all compared groups p < 0.001 ).

Cross-Polarization Optical Coherence Tomography for Brain Tumor Imaging.

This paper considers valuable visual assessment criteria for distinguishing between tumorous and non-tumorous tissues, intraoperatively, using cross-polarization OCT (CP OCT)-OCT with a functional extension, that enables detection of the polarization properties of the tissues in addition to their conventional light scattering. Materials and Methods: The study was performed on 176 ex vivo human specimens obtained from 30 glioma patients. To measure the degree to which the typical parameters of CP OCT images can be matched to the actual histology, 100 images of tumors and white matter were selected for visual analysis to be undertaken by three "single-blinded" investigators. An evaluation of the inter-rater reliability between the investigators was performed. Application of the identified visual CP OCT criteria for intraoperative use was performed during brain tumor resection in 17 patients. Results: The CP OCT image parameters that can typically be used for visual assessment were separated: (1) signal intensity; (2) homogeneity of intensity; (3) attenuation rate; (4) uniformity of attenuation. The degree of match between the CP OCT images and the histology of the specimens was significant for the parameters "signal intensity" in both polarizations, and "homogeneity of intensity" as well as the "uniformity of attenuation" in co-polarization. A test based on the identified criteria showed a diagnostic accuracy of 87-88%. Intraoperative in vivo CP OCT images of white matter and tumors have similar signals to ex vivo ones, whereas the cortex in vivo is characterized by indicative vertical striations arising from the "shadows" of the blood vessels; these are not seen in ex vivo images or in the case of tumor invasion. Conclusion: Visual assessment of CP OCT images enables tumorous and non-tumorous tissues to be distinguished. The most powerful aspect of CP OCT images that can be used as a criterion for differentiation between tumorous tissue and white matter is the signal intensity. In distinguishing white matter from tumors the diagnostic accuracy using the identified visual CP OCT criteria was 87-88%. As the CP OCT data is easily associated with intraoperative neurophysiological and neuronavigation findings this can provide valuable complementary information for the neurosurgeon tumor resection.

Quantitative nontumorous and tumorous human brain tissue assessment using microstructural co- and cross-polarized optical coherence tomography.

Optical coherence tomography (OCT) is a promising method for detecting cancer margins during tumor resection. This study focused on differentiating tumorous from nontumorous tissues in human brain tissues using cross-polarization OCT (CP OCT). The study was performed on fresh ex vivo human brain tissues from 30 patients with high- and low-grade gliomas. Different tissue types that neurosurgeons should clearly distinguish during surgery, such as the cortex, white matter, necrosis and tumorous tissue, were separately analyzed. Based on volumetric CP OCT data, tumorous and normal brain tissue were differentiated using two optical coefficients - attenuation and forward cross-scattering. Compared with white matter, tumorous tissue without necrotic areas had significantly lower optical attenuation and forward cross-scattering values. The presence of particular morphological patterns, such as necrosis and injured myelinated fibers, can lead to dramatic changes in coefficient values and create some difficulties in differentiating between tissues. Color-coded CP OCT maps based on optical coefficients provided a visual assessment of the tissue. This study demonstrated the high translational potential of CP OCT in differentiating tumorous tissue from white matter. The clinical use of CP OCT during surgery in patients with gliomas could increase the extent of tumor resection and improve overall and progression-free survival.

Intrinsic blinking of red fluorescent proteins for super-resolution microscopy.

Single-molecule localization microscopy relies on either controllable photoswitching of fluorescent probes or their robust blinking. We have found that blinking of monomeric red fluorescent proteins TagRFP, TagRFP-T, and FusionRed occurs at moderate illumination power and matches well with camera acquisition speed. It allows for super-resolution image reconstruction of densely labelled structures in live cells using various algorithms.

Quantitative evaluation of atherosclerotic plaques using cross-polarization optical coherence tomography, nonlinear, and atomic force microscopy.

A combination of approaches to the image analysis in cross-polarization optical coherence tomography (CP OCT) and high-resolution imaging by nonlinear microscopy and atomic force microscopy (AFM) at the different stages of atherosclerotic plaque development is studied. This combination allowed us to qualitatively and quantitatively assess the disorganization of collagen in the atherosclerotic arterial tissue (reduction and increase of CP backscatter), at the fiber (change of the geometric distribution of fibers in the second-harmonic generation microscopy images) and fibrillar (violation of packing and different nature of a basket-weave network of fibrils in the AFM images) organization levels. The calculated CP channel-related parameters are shown to have a statistically significant difference between stable and unstable (also called vulnerable) plaques, and hence, CP OCT could be a potentially powerful, minimally invasive method for vulnerable plaques detection.

Multi-modal optical imaging characterization of atherosclerotic plaques.

We combined cross-polarization optical coherence tomography (CP OCT) and non-linear microscopy based on second harmonic generation (SHG) and two-photon-excited fluorescence (2PEF) to assess collagen and elastin fibers and other vascular structures in the development of atherosclerosis, including identification of vulnerable plaques, which remains an important clinical problem and imaging application. CP OCT's ability to visualize tissue birefringence and cross-scattering adds new information about the microstructure and composition of the plaque. However its interpretation can be ambiguous, because backscattering contrast may have a similar appearance to the birefringence related fringes. Our results represent a step towards minimally invasive characterization and monitoring of different stages of atherosclerosis, including vulnerable plaques. CP OCT image of intimal thickening in the human coronary artery. The dark stripe in the cross-polarization channel (arrow) is a polarization fringe related to the phase retardation between two eigen polarization states. It is histologically located in the area of the lipid pool, however this stripe is a polarization artifact, rather than direct visualization of the lipid pool.

Hybrid M-mode-like OCT imaging of three-dimensional microvasculature in vivo using reference-free processing of complex valued B-scans.

We propose a novel OCT-based method for visualizing microvasculature in three-dimension using reference-free processing of individual complex valued B-scans with highly overlapped A-scans. In the lateral direction of such a B-scan, the amplitude and phase of speckles corresponding to vessel regions exhibit faster variability and, thus, can be detected without comparison with other B-scans recorded in the same plane. This method combines elements of several existing OCT angiographic approaches and exhibits: (1) enhanced robustness with respect to bulk tissue motion with frequencies up to tens of Hz, (2) resolution of microcirculation images equal to that of structural images, and (3) possibility of quantifying the vessels in terms of their decorrelation rates.