Analysis and optimization of aberration induced by oblique incidence for in-vivo tissue polarimetry.

For in-vivo polarimetry such as Mueller matrix endoscopy of human internal organ cavities, the complicated undulating tissue surfaces deliver an inescapable occurrence of oblique incidence, which induce a prominent aberration to backscattering tissue polarimetry. In this Letter, we quantitatively analyze such polarimetric aberration on polarization basic parameters derived from the Mueller matrix. A correlation heatmap is obtained as applicable criteria to select an appropriate incident angle for different polarization basic parameters. Based on the analyzing results, we propose two aberration optimization strategies of parameter selection and azimuth rotation, which are suitable for tissue samples with randomly and well-aligned fiber textures, respectively. Both strategies are demonstrated to be effective in the ex-vivo human gastric muscularis tissue experiment. The findings presented in this Letter can be useful to provide accurate polarization imaging results, widely applied on in-vivo polarimetric endoscopy for tissues with complicated surface topography.

Influence of hematoxylin and eosin staining on linear birefringence measurement of fibrous tissue structures in polarization microscopy.

Significance: For microscopic polarization imaging of tissue slices, two types of samples are often prepared: one unstained tissue section for polarization imaging to avoid possible influence from staining dyes quantitatively and one hematoxylin-eosin (H&E) stained adjacent tissue section for histological diagnosis and structural feature identification. However, this sample preparation strategy requires high-quality adjacent tissue sections, and labeling the structural features on unstained tissue sections is impossible. With the fast development of data driven-based polarimetric analysis, which requires a large amount of pixel labeled images, a possible method is to directly use H&E stained slices, which are standard samples archived in clinical hospitals for polarization measurement. Aim: We aim to study the influence of hematoxylin and eosin staining on the linear birefringence measurement of fibrous tissue structures. Approach: We examine the linear birefringence properties of four pieces of adjacent bone tissue slices with abundant collagen fibers that are unstained, H&E stained, hematoxylin (H) stained, and eosin (E) stained. After obtaining the spatial maps of linear retardance values for the four tissue samples, we carry out a comparative study using a frequency distribution histogram and similarity analysis based on the Bhattacharyya coefficient to investigate how H&E staining affects the linear birefringence measurement of bone tissues. Results: Linear retardance increased after H&E, H, or E staining (41.7%, 40.8%, and 72.5% increase, respectively). However, there is no significant change in the imaging contrast of linear retardance in bone tissues. Conclusions: The linear retardance values induced by birefringent collagen fibers can be enhanced after H&E, H, or E staining. However, the structural imaging contrasts based on linear retardance did not change significantly or the staining did not generate linear birefringence on the sample area without collagen. Therefore, it can be acceptable to prepare H&E stained slices for clinical applications of polarimetry based on such a mapping relationship.

Analyzing the influence of oblique incidence on quantitative backscattering tissue polarimetry: a pilot ex vivo study.

Significance: Among the available polarimetric techniques, backscattering Mueller matrix (MM) polarimetry provides a promising non-contact and quantitative tool for in vivo tissue detection and clinical diagnosis. To eliminate the surface reflection from the sample cost-effectively, the non-collinear backscattering MM imaging setup always has an oblique incidence. Meanwhile, for practical organ cavities imaged using polarimetric gastrointestinal endoscopy, the uneven tissue surfaces can induce various relative oblique incidences inevitably, which can affect the polarimetry in a complicated manner and needs to be considered for detailed study. Aim: The purpose of this study is to systematically analyze the influence of oblique incidence on backscattering tissue polarimetry. Approach: We measured the MMs of experimental phantom and ex vivo tissues with different incident angles and adopted a Monte Carlo simulation program based on cylindrical scattering model for further verification and analysis. Meanwhile, the results were quantitatively evaluated using the Fourier transform, basic statistics, and frequency distribution histograms. Results: Oblique incidence can induce different changes on non-periodic, two-periodic, and four-periodic MM elements, leading to false-positive and false-negative polarization information for tissue polarimetry. Moreover, a prominent oblique incidence can bring more dramatic signal variations, such as phase retardance and element transposition. Conclusions: The findings presented in this study give some crucial criterions of appropriate incident angle selections for in vivo polarimetric endoscopy and other applications and can also be valuable references for studying how to minimize the influence further.

Analyzing the Influence of Imaging Resolution on Polarization Properties of Scattering Media Obtained From Mueller Matrix.

The Mueller matrix contains abundant micro- and even nanostructural information of media. Especially, it can be used as a powerful tool to characterize anisotropic structures quantitatively, such as the particle size, density, and orientation information of fibers in the sample. Compared with unpolarized microscopic imaging techniques, Mueller matrix microscopy can also obtain some essential structural information about the sample from the derived parameters images at low resolution. Here, to analyze the comprehensive effects of imaging resolution on polarization properties obtained from the Mueller matrix, we, first, measure the microscopic Mueller matrices of unstained rat dorsal skin tissue slices rich in collagen fibers using a series of magnifications or numerical aperture (NA) values of objectives. Then, the first-order moments and image texture parameters are quantified and analyzed in conjunction with the polarization parameter images. The results show that the Mueller matrix polar decomposition parameters diattenuation D, linear retardance δ, and depolarization Δ images obtained using low NA objective retain most of the structural information of the sample and can provide fast imaging speed. In addition, the scattering phase function analysis and Monte Carlo simulation based on the cylindrical scatterers reveal that the diattenuation parameter D images with different imaging resolutions are expected to be used to distinguish among the fibrous scatterers in the medium with different particle sizes. This study provides a criterion to decide which structural information can be accurately and rapidly obtained using a transmission Mueller matrix microscope with low NA objectives to assist pathological diagnosis and other applications.

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.