Cross-polarized Imaging Research Articles

Noninvasive assessment of cutaneous alterations in mice exposed to whole body gamma irradiation using optical imaging techniques.

We report the results of a study carried out to investigate the potential of optical techniques such as optical coherence tomography, Mueller matrix spectroscopy, and cross-polarization imaging for noninvasive monitoring of the ionizing radiation exposure-induced alterations in cutaneous tissue of mice. Radiation dose-dependent changes were observed in tissue microvasculature and tissue optical parameters like retardance and depolarization as early as 1h post radiation exposure. Results suggest that these optical techniques may allow early detection of radiation dose-dependent alterations which could help in screening of population exposed to radiation.

Tensile Mechanical Properties and Dynamic Collagen Fiber Re-Alignment of the Murine Cervix are Dramatically Altered Throughout Pregnancy.

The cervix is a unique organ able to dramatically change its shape and function by serving as a physical barrier for the growing fetus and then undergoing dramatic dilation allowing for delivery of a term infant. As a result, the cervix endures changing mechanical forces from the growing fetus. There is an emerging concept that the cervix may change or remodel "early" in many cases of spontaneous preterm birth (sPTB). However, the mechanical role of the cervix in both normal and preterm birth remains unclear. Therefore, the primary objective of this study was to determine the mechanical and structural responses of murine cervical tissue throughout a normal gestational time course. In this study, both tissue structural and material properties were determined via a quasi-static tensile load-to-failure test, while simultaneously obtaining dynamic collagen fiber re-alignment via cross-polarization imaging. This study demonstrated that the majority of the mechanical properties evaluated decreased at midgestation and not just at term, while collagen fiber re-alignment occurred earlier in the loading curve for cervices at term. This suggests that although structural changes in the cervix occur throughout gestation, the differences in material properties function in combination with collagen fiber re-alignment as mechanical precursors to regulate term gestation. This work lays a foundation for investigating cervical biomechanics and the role of the cervix in preterm birth.

Diversity in the organization of elastin bundles and intramembranous muscles in bat wings

Unlike birds and insects, bats fly with wings composed of thin skin that envelops the bones of the forelimb and spans the area between the limbs, digits, and sometimes the tail. This skin is complex and unusual; it is thinner than typical mammalian skin and contains organized bundles of elastin and embedded skeletal muscles. These elements are likely responsible for controlling the shape of the wing during flight and contributing to the aerodynamic capabilities of bats. We examined the arrangement of two macroscopic architectural elements in bat wings, elastin bundles and wing membrane muscles, to assess the diversity in bat wing skin morphology. We characterized the plagiopatagium and dactylopatagium of 130 species from 17 families of bats using cross-polarized light imaging. This method revealed structures with distinctive relative birefringence, heterogeneity of birefringence, variation in size, and degree of branching. We used previously published anatomical studies and tissue histology to identify birefringent structures, and we analyzed their architecture across taxa. Elastin bundles, muscles, neurovasculature, and collagenous fibers are present in all species. Elastin bundles are oriented in a predominantly spanwise or proximodistal direction, and there are five characteristic muscle arrays that occur within the plagiopatagium, far more muscle than typically recognized. These results inform recent functional studies of wing membrane architecture, support the functional hypothesis that elastin bundles aid wing folding and unfolding, and further suggest that all bats may use these architectural elements for flight. All species also possess numerous muscles within the wing membrane, but the architecture of muscle arrays within the plagiopatagium varies among families. To facilitate present and future discussion of these muscle arrays, we refine wing membrane muscle nomenclature in a manner that reflects this morphological diversity. The architecture of the constituents of the skin of the wing likely plays a key role in shaping wings during flight.

Direct deconvolution of electric and magnetic responses of single nanoparticles by Fourier space surface plasmon resonance microscopy

We use polarization-resolved surface plasmon resonance microscopy to image single dielectric nanoparticles. In real space, the nanoparticles exhibit V-shape diffraction patterns due to the interference between the incident surface plasmon polariton wave and the evanescent scattered waves, which arise from the interplay between the electric and magnetic dipoles of the nanoparticle. By using cross-polarized Fourier space imaging to extract only the scattered waves, we find the angular far-field intensity corresponds very well to the near-field scattering distribution, as confirmed by both analytical and numerical calculations. As a result, we directly deconvolute the contributions of electric and magnetic dipoles to the scattered fields without involving near-field techniques.

Cross-polarized GPR imaging of fracture flow channeling

Ground penetrating radar (GPR) can be used to image fractures and monitor fluid flow in the subsurface. Conventional GPR imaging uses single-polarization, co-polarized acquisition. We examine the use of cross-polarized GPR signals for imaging flow channeling in a discrete horizontal fracture. Numerical modeling (FDTD) demonstrates that when the fracture channel is oriented at an oblique angle to the survey line, depolarization of the GPR signal results in scattered energy along the cross-polarized components. When the channel is oriented parallel or orthogonal to the survey line, all scattered energy is captured by the co-polarized components and no signal is present in the cross-polarized orientation. Multipolarization, time-lapse 3D GPR field data were acquired at the Altona Flat Rock test site in New York State. The GPR surveys were conducted during background fresh fracture water conditions and during a natural gradient saline tracer test which was used to highlight flow channels along a sub-horizontal fracture. Amplitude analysis of the cross-polarized data reveals flow channeling that is in agreement with the co-polarized GPR images and with independent hydraulic tests. This investigation demonstrates that cross-polarized components of GPR signals can be used to enhance imaging of flow channels in fractured media.

Flow-through cross-polarized imaging as a new tool to overcome the analytical sensitivity challenges of a low-dose crystalline compound in a lipid matrix

Assessing the physical state of a low-dose active compound in a solid lipid or polymer matrix is analytically challenging, especially if the matrix exhibits some crystallinity. The aim of this study was first to compare the ability of current methods to detect the presence of a crystalline model compound in lipid matrices. Subsequently, a new technique was introduced and evaluated because of sensitivity issues that were encountered with current methods. The new technique is a flow-through version of cross-polarized imaging in transmission mode. The tested lipid-based solid dispersions (SDs) consisted of β-carotene (BC) as a model compound, and of Gelucire 50/13 or Geleol mono- and diglycerides as lipid matrices. The solid dispersions were analyzed by (hyper) differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), and microscopic techniques including atomic force microscopy (AFM). DSC and XRPD could analyze crystalline BC at concentrations as low as 3% (w/w) in the formulations. However, with microscopic techniques crystalline particles were detected at significantly lower concentrations of even 0.5% (w/w) BC. A flow-through cross-polarized imaging technique was introduced that combines the advantage of analyzing a larger sample size with high sensitivity of microscopy. Crystals were detected easily in samples containing even less than 0.2% (w/w) BC. Moreover, the new tool enabled approximation of the kinetic BC solubility in the crystalline lipid matrices. As a conclusion, the flow-through cross-polarized imaging technique has the potential to become an indispensable tool for characterizing low-dose crystalline compounds in a lipid or polymer matrix of solid dispersions.

Wavelength optimized cross-polarized wide-field imaging for noninvasive and rapid evaluation of dermal structures.

Changes in the morphology of dermal collagen may indicate aging or pathological processes. At present, there is no technology for in vivo real-time assessment of collagen structures. Our goal was to introduce and validate polarization optical imaging for noninvasive quantitative evaluation of dermal collagen. Seventeen volunteers participated in the study. Cross-polarized 440nm images were acquired noninvasively from facial skin of the study subjects. Collagen content and intensity histogram were computed from the optical images. Quantitative results showed a decrease in the collagen content with increasing age of the subjects. Analysis of the collagen image histogram parameters demonstrated decreasing mean pixel value and increasing full width at half maximum (FWHM) with increasing age. Polarization optical imaging has the potential for rapid noninvasive in vivo evaluation of human dermis. Cross-polarized imaging at different wavelengths emphasizes different features of human skin. (A) Skin photograph. (B) In vivo cross-polarized image at 440 nm. (C) In vivo cross-polarized image at 570nm. (D) In vivo cross-polarized image at 690nm.

Modular video endoscopy forin vivocross-polarized and vital-dye fluorescence imaging of Barrett’s-associated neoplasia

A modular video endoscope is developed and tested to allow imaging in different modalities. This system incorporates white light imaging (WLI), cross-polarized imaging (CPI), and vital-dye fluorescence imaging (VFI), using interchangeable filter modules. CPI and VFI are novel endoscopic modalities that probe mucosal features associated with Barrett's neoplasia. CPI enhances vasculature, while VFI enhances glandular architecture. In this pilot study, we demonstrate the integration of these modalities by imaging areas of Barrett's metaplasia and neoplasia in an esophagectomy specimen. We verify that those key image features are also observed during an in vivo surveillance procedure. CPI images demonstrate improved visualization of branching blood vessels associated with neoplasia. VFI images show glandular architecture with increased glandular effacement associated with neoplasia. Results suggests that important pathologic features seen in CPI and VFI are not visible during standard endoscopic white light imaging, and thus the modalities may be useful in future in vivo studies for discriminating neoplasia from Barrett's metaplasia. We further demonstrate that the integrated WLI/CPI/VFI endoscope is compatible with complementary high-resolution endomicroscopy techniques such as the high-resolution microendoscope, potentially enabling two-step ("red-flag" widefield plus confirmatory high-resolution imaging) protocols to be enhanced.

新型全光纤交叉偏振光学相干层析成像系统

新型全光纤交叉偏振光学相干层析成像系统

Noninvasive clinical assessment of port‐wine stain birthmarks using current and future optical imaging technology: a review

Port-wine stain (PWS) birthmarks are one class of benign congenital vascular malformation. Laser therapy is the most successful treatment modality of PWS. Unfortunately, this approach has limited efficacy, with only 10% of patients experiencing complete blanching of the PWS. To address this problem, several research groups have developed technologies and methods designed to study treatment outcome and improve treatment efficacy. This article reviews seven optical imaging techniques currently in use or under development to assess treatment efficacy, focusing on: reflectance spectrophotometers/tristimulus colorimeters; laser Doppler flowmetry and laser Doppler imaging; cross-polarized diffuse reflectance colour imaging system; reflectance confocal microscopy; optical coherence tomography; spatial frequency domain imaging; and laser speckle imaging.

Characterization of Semi-Insulating 4H Silicon Carbide for Radiation Detectors

Radiation detectors have been fabricated on 8 mm × 8 mm substrates, ~390 μm in thickness, diced from a (0001) 4H-SiC semi-insulating (SI) wafer (≥ 1012 Ohm-cm). The crystals used for detector fabrication have been characterized by x-ray diffraction (XRD), electron beam induced current (EBIC), chemical etching, cross-polarized imaging, and Raman spectroscopy. Current-voltage (I-V) characteristics showed very low leakage current (≤ 50 pA at -800 V) and the capability of detector's operation ≥ 470 K. EBIC investigations revealed that the screw dislocations produce dark EBIC contrast indicating high leakage current in the defective regions. Thermally stimulated current (TSC) measurements and high temperature resistivity measurements revealed deep level centers with activation energies 1.1-1.2 eV, and 1.56 eV. The TSC peak at ~460 K associated with the ~1.2 eV center was much stronger than the other high temperature peaks (e.g., 370 K due to vanadium impurity, 0.95 eV below of conduction band edge), indicating that this level along with the 1.56 eV level should dominate in controlling the resistivity and carrier lifetime in the studied 4H-SiC. Based on the literature data, we associate these centers with intrinsic defects and/or V-related complex. Nuclear detection measurements on the single-element SiC detectors with 241Am X-γ ray source clearly detected 59.6 keV and other low energy x-rays.

Detection of malarial byproduct hemozoin utilizing its unique scattering properties

The scattering characteristics of the malaria byproduct hemozoin, including its scattering distribution and depolarization, are modeled using Discrete Dipole Approximation (DDA) and compared to those of healthy red blood cells. Scattering (or dark-field) spectroscopy and imaging are used to identify hemozoin in fresh rodent blood samples. A new detection method is proposed and demonstrated using dark-field in conjunction with cross-polarization imaging and spectroscopy. SNRs greater than 50:1 are achieved for hemozoin in fresh blood without the addition of stains or reagents. The potential of such a detection system is discussed.

Wide-field and high-resolution optical imaging for early detection of oral neoplasia

Current procedures for oral cancer screening typically involve visual inspection of the entire tissue surface at risk under white light illumination. However, pre-cancerous lesions can be difficult to distinguish from many benign conditions when viewed under these conditions. We have developed wide-field (macroscopic) imaging system which additionally images in cross-polarized white light, narrowband reflectance, and fluorescence imaging modes to reduce specular glare, enhance vascular contrast, and detect disease-related alterations in tissue autofluorescence. We have also developed a portable system to enable high-resolution (microscopic) evaluation of cellular features within the oral mucosa in situ. This system is a wide-field epi-fluorescence microscope coupled to a 1 mm diameter, flexible fiber-optic imaging bundle. Proflavine solution was used to specifically label cell nuclei, enabling the characteristic differences in N/C ratio and nuclear distribution between normal, dysplastic, and cancerous oral mucosa to be quantified. This paper discusses the technical design and performance characteristics of these complementary imaging systems. We will also present data from ongoing clinical studies aimed at evaluating diagnostic performance of these systems for detection of oral neoplasia.

Polarization microscopy with stellated gold nanoparticles for robust, in-situ monitoring of biomolecules

Advances in plasmonic nanoparticle synthesis afford new opportunities for biosensing applications. Here, we apply a combination of a new type of plasmonic nanomaterial - stellated nanoparticles, and polarization-sensitive darkfield microscopy for detecting molecular assemblies and tracking of individual epidermal growth factor receptors within single live cells with high signal-to-background ratio. Depolarization of linear polarized light by stellated nanoparticles is over 15-fold more efficient than similarly-sized spheroidal nanoparticles. This efficient light depolarization allows robust detection of molecules labeled with stellated nanoparticles in cross-polarized imaging where the intrinsic light scattering from cells is significantly reduced. The imaging can be carried out with single molecule sensitivity for essentially unlimited time with no signal degradation.

Near-infrared imaging of fast intrinsic optical responses in visible light-activated amphibian retina

High performance functional imaging is needed for dynamic measurements of neural processing in retina. Emerging techniques for visual prosthesis also require advanced methodology for reliable validation of electromagnetic stimulation of the retina. Imaging of fast intrinsic optical responses associated with neural activation promises a variety of technical advantages over traditional single and multichannel electrophysiological techniques for these purposes, but the application of fast optical signals for neural imaging has been limited by low signal-to-noise ratio and high background light intensity. However, by using an optimized near-infrared probe light and improved optical system, we improve the optical signals substantially, allowing single pass measurements with approximately micron resolution. We image fast intrinsic optical responses with different optical modalities, i.e., bright field, dark field, and cross-polarization, from isolated retina activated by visible light stimulation. At single cell resolution, bright-field imaging discloses the maxima of optical responses approximately 5% dI/I, where dI is the dynamic optical change and I is the baseline light intensity. Dark-field imaging techniques further enhance the sensitivity of optical measurements, and show the maxima of optical responses exceeding 10% dI/I. Cross-polarized imaging provides optical sensitivity similar to dark-field imaging, but different patterns of neural activation are observed.

Applying the polarization memory effect in polarization-gated subsurface imaging

Polarization memory is a well established phenomenon occurring when circularly polarized light propagates in turbid media of larger particles. Recent studies have demonstrated that the circularly cross-polarized imaging can significantly improve subsurface reflection contrast due to the polarization memory effect. We have found that such improvement is strongly influenced by the optical properties of the media. Circularly cross-polarized light provides superior image enhancement in low scattering media, but becomes inferior in high scattering media. Our experiments also demonstrate that polarization imaging provides no significant improvement to image resolution.

Spectroscopic imaging for detection of ischemic injury in rat kidneys by use of changes in intrinsic optical properties

It is currently impossible to consistently predict kidney graft viability and function before and after transplantation. We explored optical spectroscopy to assess the degree of ischemic damage in kidney tissue. Tunable UV laser excitation was used to record autofluorescence images, at different spectral ranges, of injured and contralateral control rat kidneys to reveal the excitation conditions that offered optimal contrast. Autofluorescence and near-infrared cross-polarized light-scattering imaging were both used to monitor changes in intensity and spectral characteristics, as a function of exposure time to ischemic injury. These two modalities provided different temporal behaviors, arguably arising from two different mechanisms providing direct correlation of intrinsic optical signatures to ischemic injury time.

Cross-polarization imaging and micro-raman detection of defects in the epitaxy of 4H-SiC

Cross-polarized imaging with an optical scanner and confocal μ-Raman spectroscopy have been used to reveal and to study structural defects in n-4H-SiC. We introduce a new method of polarized imaging that uses a combination of linear and circular polarization, which enhances contrast in images of defect regions. Regions that show intensity contrast have been observed to have characteristics of either low-angle grain boundaries or sharp lines of delineation between low and high strain areas. Different types of polytype inclusions have also been observed, and the material within the inclusions has been shown to be either 3C-SiC or 6H-SiC. Polytype inclusions occur as isolated features or form cores of holes, and they can form at any point of the growth.

Characterization of port wine stain skin erythema and melanin content using cross-polarized diffuse reflectance imaging.

Objective methods to assess quantitatively port wine stain (PWS) blanching in response to laser therapy are needed to improve laser therapeutic outcome. Previous studies have attempted to assess objectively PWS color based on point measurement devices. To date, these approaches have typically been limited by a number of factors such as small test area and need for contact. To address these issues, a cross-polarized diffuse reflectance imaging system and color image analysis method has been developed to evaluate quantitatively erythema and melanin content in PWS skin. A cross-polarized diffuse reflectance system has been constructed to acquire high resolution digital images while minimizing artifacts such as glare, shadowing, and nonuniform illumination effects that can compromise image fidelity. Furthermore, an image analysis algorithm has also been developed to analyze normal and PWS skin in terms of CIEL*a*b* color space parameters. Using the algorithm, images of color space intensity, L*, and saturation, a*, indices have been calculated to extract quantitative metrics of melanin and erythema, respectively. Compared to the cross-polarized diffuse reflectance image, the a* index image enhanced the contrast in regions of high and low erythema and seems to have quasi correlation with the L* index image. In the presented PWS patient example, the relative a* index difference (Deltaa*) image showed less erythema in PWS skin after laser treatment when compared to pre-treatment values. Our cross-polarized imaging system and color image analysis method is a simple, noncontact technique that can indirectly provide quantitative measurement of erythema and melanin content in PWS skin.

Calibrated fluorescence imaging of tissue in vivo

A calibrated fluorescence imaging system utilizing a combination of fluorescence and cross-polarization imaging technology is described and applied to tissue examination in vivo. The results show that the inhomogeneity of fluorescence excitation and collection across the irregular surface of the examined tissue is calibrated to a great extent by taking the ratio of the raw fluorescence image to the cross-polarized reflection image. The effects of optical properties of tissue on the calibrated fluorescence signals are studies on simulated tissue phantoms systematically. Using the calibrated fluorescence imaging technology, we demonstrate that different tissue can be clearly separated endoscopically and in vivo based on the calibrated fluorescence signal.