Polarization enhancement mechanism from tissue staining in multispectral Mueller matrix microscopy.

Mueller matrix microscopy can provide comprehensive polarization-related optical and structural information of biomedical samples label-freely. Thus, it is regarded as an emerging powerful tool for pathological diagnosis. However, the staining dyes have different optical properties and staining mechanisms, which can put influence on Mueller matrix microscopic measurement. In this Letter, we quantitatively analyze the polarization enhancement mechanism from hematoxylin and eosin (H&E) staining in multispectral Mueller matrix microscopy. We examine the influence of hematoxylin and eosin dyes on Mueller matrix-derived polarization characteristics of fibrous tissue structures. Combined with Monte Carlo simulations, we explain how the dyes enhance diattenuation and linear retardance as the illumination wavelength changed. In addition, it is demonstrated that by choosing an appropriate incident wavelength, more visual Mueller matrix polarimetric information can be observed of the H&E stained tissue sample. The findings can lay the foundation for the future Mueller matrix-assisted digital pathology.

Transmission Mueller matrix imaging with spatial filtering.

In this Letter, we report a study on the effects of spatial filtering for a transmission Mueller matrix imaging system. A spatial filter (SF) is placed on the back Fourier plane of the imaging lens in a dual-rotating-retarders Mueller matrix imaging system to select photons within a certain scattering angle. The system is then applied to three types of human cancerous tissues. When imaging with a small-aperture SF, some polarimetry basis parameters show sharp changes in contrast in the cancerous regions. Monte Carlo simulations using a simple sphere-cylinder scattering model also show that spatial filtering of the scattered photons provides extra information on the size and shape of the scattering particles. The results indicate that spatial filtering enhances the capability of polarization imaging as a powerful tool for biomedical diagnosis.

Characteristic Mueller matrices for direct assessment of the breaking of symmetries.

Mueller polarimetry is a powerful optical technique in the analysis of micro-structural properties of optical samples. However, there is no explicit relationship between individual Mueller matrix elements and the physical properties of the sample. Several matrix decomposition algorithms corresponding to specific optical models have been proposed, which extract the physical information from measured Mueller matrices. Nevertheless, we still need a prior assessment method to decide which model is more suitable for the experimental data. In this Letter, we propose a set of characteristic Mueller matrices that allows us to obtain information about the breaking of rotation, mirror, and reciprocal symmetry properties in the sample by direct inspection of several elements of the Mueller matrix. By further analyzing the possible origin of symmetry breaking, we can learn the type and mixing status of anisotropies in the measured sample. We have verified our theory with Monte Carlo simulations of polarized light scattering in an isotropic or anisotropic medium containing different configurations of spherical and cylindrical scatterers. This study may help experimenters choose more suitable Mueller matrix decomposition methods.

Assessment of tissue polarimetric properties using Stokes polarimetric imaging with circularly polarized illumination.

Tissue-depolarization and linear-retardance are the main polarization characteristics of interest for bulk tissue characterization, and are normally interpreted from Mueller polarimetry. Stokes polarimetry can be conducted using simpler instrumentation and in a shorter time. Here, we use Stokes polarimetric imaging with circularly polarized illumination to assess the circular-depolarization and linear-retardance properties of tissue. Results obtained were compared with Mueller polarimetry in transmission and reflection geometry, respectively. It is found that circular-depolarization obtained from these 2 methods is very similar in both geometries, and that linear-retardance is highly quantitatively similar for transmission geometry and qualitatively similar for reflection geometry. The majority of tissue circular-depolarization and linear-retardance image information (represented by local image contrast features) obtained from Mueller polarimetry is well preserved from Stokes polarimetry in both geometries. These findings can be referred to for further understanding tissue Stokes polarimetric data, and for further application of Stokes polarimetry under the circumstances where short acquisition time or low optical system complexity is a priority, such as polarimetric endoscopy and microscopy.

A quantitative and non-contact technique to characterise microstructural variations of skin tissues during photo-damaging process based on Mueller matrix polarimetry.

Skin tissue consists of collagen and elastic fibres, which are highly susceptible to damage when exposed to ultraviolet radiation (UVR), leading to skin aging and cancer. However, a lack of non-invasive detection methods makes determining the degree of UVR damage to skin in real time difficult. As one of the fundamental features of light, polarization can be used to develop imaging techniques capable of providing structural information about tissues. In particular, Mueller matrix polarimetry is suitable for detecting changes in collagen and elastic fibres. Here, we demonstrate a novel, quantitative, non-contact and in situ technique based on Mueller matrix polarimetry for monitoring the microstructural changes of skin tissues during UVR-induced photo-damaging. We measured the Mueller matrices of nude mouse skin samples, then analysed the transformed parameters to characterise microstructural changes during the skin photo-damaging and self-repairing processes. Comparisons between samples with and without the application of a sunscreen showed that the Mueller matrix-derived parameters are potential indicators for fibrous microstructure in skin tissues. Histological examination and Monte Carlo simulations confirmed the relationship between the Mueller matrix parameters and changes to fibrous structures. This technique paves the way for non-contact evaluation of skin structure in cosmetics and dermatological health.

Monitoring microstructural variations of fresh skeletal muscle tissues by Mueller matrix imaging.

Recently many attempts have been made for extracting the structural information of myofibrils as indicators for diseases of skeletal muscle. In this paper we adopt wide-field illumination and take the backscattering Mueller matrix images of bovine skeletal muscle tissues during the 24-hour experimental time after the animal's death. The 2D images of Mueller matrix elements and their frequency distribution histograms (FDHs) reveal rich qualitative information on the changes in the microstructures of the skeletal muscle. The temporal variations of the sample are quantitatively analyzed using two Mueller matrix transformation (MMT) parameters. The characteristic features of the temporal plots are attributed to the rigor mortis and proteolysis processes. For a deeper insight on the relationship between the features of the MMT parameters and the microstructures during the rigor mortis and proteolysis processes, Monte Carlo (MC) simulations are carried out based on sphere-cylinder birefringence model (SCBM). The good agreement between the experimental and MC simulated results show that the FDHs and MMT parameters can describe more clearly the characteristic microstructural features of skeletal muscle tissues. The techniques are useful for the characterization of physiological status of tissues, or quantitative assessment of meat qualities in food industry.

GPU acceleration of Monte Carlo simulations for polarized photon scattering in anisotropic turbid media.

In earlier studies, we developed scattering models and the corresponding CPU-based Monte Carlo simulation programs to study the behavior of polarized photons as they propagate through complex biological tissues. Studying the simulation results in high degrees of freedom that created a demand for massive simulation tasks. In this paper, we report a parallel implementation of the simulation program based on the compute unified device architecture running on a graphics processing unit (GPU). Different schemes for sphere-only simulations and sphere-cylinder mixture simulations were developed. Diverse optimizing methods were employed to achieve the best acceleration. The final-version GPU program is hundreds of times faster than the CPU version. Dependence of the performance on input parameters and precision were also studied. It is shown that using single precision in the GPU simulations results in very limited losses in accuracy. Consumer-level graphics cards, even those in laptop computers, are more cost-effective than scientific graphics cards for single-precision computation.

Characterizing the Effects of Washing by Different Detergents on the Wavelength-Scale Microstructures of Silk Samples Using Mueller Matrix Polarimetry.

Silk fibers suffer from microstructural changes due to various external environmental conditions including daily washings. In this paper, we take the backscattering Mueller matrix images of silk samples for non-destructive and real-time quantitative characterization of the wavelength-scale microstructure and examination of the effects of washing by different detergents. The 2D images of the 16 Mueller matrix elements are reduced to the frequency distribution histograms (FDHs) whose central moments reveal the dominant structural features of the silk fibers. A group of new parameters are also proposed to characterize the wavelength-scale microstructural changes of the silk samples during the washing processes. Monte Carlo (MC) simulations are carried out to better understand how the Mueller matrix parameters are related to the wavelength-scale microstructure of silk fibers. The good agreement between experiments and simulations indicates that the Mueller matrix polarimetry and FDH based parameters can be used to quantitatively detect the wavelength-scale microstructural features of silk fibers. Mueller matrix polarimetry may be used as a powerful tool for non-destructive and in situ characterization of the wavelength-scale microstructures of silk based materials.

Retardance of bilayer anisotropic samples consisting of well-aligned cylindrical scatterers and birefringent media.

Both cylindrical scatterers and birefringent media may contribute to the anisotropy of tissue, where anisotropy can be characterized using polarization techniques. Our previous studies have shown that a layer of well-aligned cylindrical scatterers displays anisotropic properties similar to those of a piece of birefringent media, whose equivalent extraordinary axis is along the axial direction of the cylinders. We focused on a sample consisting of two layers of anisotropic media, with each layer having a different orientation; the characteristics of this sample were representative of the properties of multilayer fibrous tissues. Using a Mueller matrix decomposition method, we examined in detail how the total retardance and the equivalent extraordinary axis of the bilayered sample varied with changes in the retardance of the two layers and the direction of the extraordinary axis. The results of this study showed that, in such bilayer samples, a layer of well-aligned cylindrical scatterers generated a retardance that behaved exactly like the retardance generated by a piece of birefringent media. The simulated results were also confirmed by the results of experiments using aligned glass fibers.

Study of optical clearing in polarization measurements by Monte Carlo simulations with anisotropic tissue-mimicking models.

We conducted Monte Carlo simulations based on anisotropic sclera-mimicking models to examine the polarization features in Mueller matrix polar decomposition (MMPD) parameters during the refractive index matching process, which is one of the major mechanisms of optical clearing. In a preliminary attempt, by changing the parameters of the models, wavelengths, and detection geometries, we demonstrate how the depolarization coefficient and retardance vary during the refractive index matching process and explain the polarization features using the average value and standard deviation of scattering numbers of the detected photons. We also study the depth-resolved polarization features during the gradual progression of the refractive index matching process. The results above indicate that the refractive index matching process increases the depth of polarization measurements and may lead to higher contrast between tissues of different anisotropies in deeper layers. MMPD-derived polarization parameters can characterize the refractive index matching process qualitatively.

Mueller matrix microscope: a quantitative tool to facilitate detections and fibrosis scorings of liver cirrhosis and cancer tissues.

Today the increasing cancer incidence rate is becoming one of the biggest threats to human health.Among all types of cancers, liver cancer ranks in the top five in both frequency and mortality rate all over the world. During the development of liver cancer, fibrosis often evolves as part of a healing process in response to liver damage, resulting in cirrhosis of liver tissues. In a previous study, we applied the Mueller matrix microscope to pathological liver tissue samples and found that both the Mueller matrix polar decomposition (MMPD) and Mueller matrix transformation (MMT) parameters are closely related to the fibrous microstructures. In this paper,we take this one step further to quantitatively facilitate the fibrosis detections and scorings of pathological liver tissue samples in different stages from cirrhosis to cancer using the Mueller matrix microscope. The experimental results of MMPD and MMT parameters for the fibrotic liver tissue samples in different stages are measured and analyzed. We also conduct Monte Carlo simulations based on the sphere birefringence model to examine in detail the influence of structural changes in different fibrosis stages on the imaging parameters. Both the experimental and simulated results indicate that the polarized light microscope and transformed Mueller matrix parameter scan provide additional quantitative information helpful for fibrosis detections and scorings of liver cirrhosis and cancers. Therefore, the polarized light microscope and transformed Mueller matrix parameters have a good application prospect in liver cancer diagnosis.

Study on the validity of 3 × 3 Mueller matrix decomposition.

Using Monte Carlo simulations based on previously developed scattering models consisting of spherical and cylindrical scatterers imbedded in birefringent interstitial medium, we compare the polarization parameters extracted from the 3×3 and 4×4 Mueller matrix decomposition methods in forward and backward scattering directions. The results show that the parameters derived from the 3×3 Mueller matrix decomposition are usually not the same as those from the 4×4 Mueller matrix decomposition but display similar qualitative relations to changes in the microstructure of the sample, such as the density, size, and orientation distributions of the scatterers, and birefringence of the interstitial medium. The simulations are backed up by experiments when suitable samples are available.

Mapping local orientation of aligned fibrous scatterers for cancerous tissues using backscattering Mueller matrix imaging.

Polarization measurements are sensitive to the microstructure of tissues and can be used to detect pathological changes. Many tissues contain anisotropic fibrous structures. We obtain the local orientation of aligned fibrous scatterers using different groups of the backscattering Mueller matrix elements. Experiments on concentrically well-aligned silk fibers and unstained human papillary thyroid carcinoma tissues show that the m22 , m33 , m23 , and m32 elements have better contrast but higher degeneracy for the extraction of orientation angles. The m12 and m13 elements show lower contrast, but allow us to determine the orientation angle for the fibrous scatterers along all directions. Moreover, Monte Carlo simulations based on the sphere-cylinder scattering model indicate that the oblique incidence of the illumination beam introduces some errors in the orientation angles obtained by both methods. Mapping the local orientation of anisotropic tissues may not only provide information on pathological changes, but can also give new leads to reduce the orientation dependence of polarization measurements.

Mueller matrix polarimetry for differentiating characteristic features of cancerous tissues.

Polarization measurements allow one to enhance the imaging contrast of superficial tissues and obtain new polarization sensitive parameters for better descriptions of the micro- and macro- structural and optical properties of complex tissues. Since the majority of cancers originate in the epithelial layer, probing the morphological and pathological changes in the superficial tissues using an expended parameter set with improved contrast will assist in early clinical detection of cancers. We carry out Mueller matrix imaging on different cancerous tissues to look for cancer specific features. Using proper scattering models and Monte Carlo simulations, we examine the relationship between the microstructures of the samples, which are represented by the parameters of the scattering model and the characteristic features of the Mueller matrix. This study gives new clues on the contrast mechanisms of polarization sensitive measurements for different cancers and may provide new diagnostic techniques for clinical applications.

Study on retardance due to well-ordered birefringent cylinders in anisotropic scattering media.

We report an anisotropic tissue model containing well-ordered birefringent cylinders. Using simulations and experiments, we examined the different polarization features for nonbirefringent and birefringent cylinders and analyzed the influence of the birefringent cylinders on the retardance obtained from Mueller matrix polar decomposition. For the well-ordered birefringent cylinders, retardance increases linearly with the intrinsic birefringence and the scattering coefficient. Furthermore, the cylinders with a larger diameter generate more retardance. Compared with the cylinder-birefringence model, in which birefringent medium exists between the scatterers, the intrinsic birefringence on the cylinders usually contributes much less to the total retardance.

Two-dimensional and surface backscattering Mueller matrices of anisotropic sphere-cylinder scattering media: a quantitative study of influence from fibrous scatterers.

We present both the two-dimensional backscattering point-illumination and surface-illumination Mueller matrices for the anisotropic sphere-cylinder scattering media. The experimental results of the microsphere-silk sample show that the Mueller matrix elements of an anisotropic scattering medium are different from those of an isotropic medium. Moreover, both the experiments and Monte Carlo simulations show that the directions of the fibrous scatterers have prominent effects on the Mueller matrix elements. As the fibrous samples rotate, the surface-illumination Mueller matrix measurement results for the m12, m21, m13, m31, m22, m23, m32, and m33 elements represent periodical variations. Experiments on skeletal muscle and porcine liver tissue samples confirm that the periodical changes for the surface-illumination Mueller matrix elements are closely related to the well aligned fibrous scatterers. The m22, m23, m32, and m33 elements are powerful tools for quantitative characterization of anisotropic scattering media, including biological tissues.

Two-dimensional backscattering Mueller matrix of sphere-cylinder birefringence media.

We have developed a sphere-cylinder birefringence model (SCBM) for anisotropic media. The new model is based on a previously published sphere-cylinder scattering model (SCSM), but the spherical and cylindrical scatterers are embedded in a linearly birefringent medium. A Monte Carlo simulation program for SCBM was also developed by adding a new module to the SCSM program to take into account the effects of birefringence. Simulations of the backscattering Mueller matrix demonstrate that SCBM results in better agreement with experimental results than SCSM and is more suitable to characterize fibrous tissues such as skeletal muscle. Using Monte Carlo simulations, we also examined the characteristics of two-dimensional backscattering Mueller matrix of SCBM and analyzed the influence of linear birefringence.

Application of sphere-cylinder scattering model to skeletal muscle.

By comparing the spatially resolved unpolarized, polarized reflectance and Mueller matrix elements of skeletal muscle with a scattering medium containing polystyrene microspheres and silk fibers, we demonstrate that the sphere-cylinder scattering model (SCSM) can reproduce the characteristic features of skeletal muscle. Both experiments and polarization sensitive Monte Carlo simulation provide evidences that SCSM may be used to characterize the structural and optical properties of skeletal muscle.

Two-dimensional backscattering Mueller matrix of sphere-cylinder scattering medium.

We present the experimental results for the two-dimensional backscattering Mueller matrix of a scattering medium containing polystyrene microspheres and silk fibers and simulate the same Mueller matrix using a polarization-sensitive Monte Carlo program with both layered and homogeneous sphere-cylinder scattering models. We discuss the characteristic features in each Mueller matrix element and their relations with the parameters of the spherical and cylindrical scatterers in the medium. Both experiments and simulations suggest that the Mueller matrix elements can be used to characterize the structural and optical properties of anisotropic scattering media.