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Significance : Definitive diagnostics of many diseases is based on the histological analysis of thin tissue cuts with optical white light microscopy. Extra information on tissue structural properties obtained with polarized light would help the pathologist to improve the accuracy of his diagnosis. Aim: We report on using Mueller matrix microscopy data, logarithmic decomposition, and polarized Monte Carlo (MC) modeling for qualitative and quantitative analysis of thin tissue cuts to extract the information on tissue microstructure that is not available with a conventional white light microscopy. Approach: Unstained cuts of human skin equivalents were measured with a custom-built liquid-crystal-based Mueller microscope in transmission configuration. To interpret experimental data, we performed the simulations with a polarized MC algorithm for scattering anisotropic media. Several optical models of tissue (spherical scatterers within birefringent host medium, and combination of spherical and cylindrical scatterers within either isotropic or birefringent host medium) were tested. Results: A set of rotation invariants for the logarithmic decomposition of a Mueller matrix was derived to rule out the impact of sample orientation. These invariants were calculated for both simulated and measured Mueller matrices of the dermal layer of skin equivalents. We demonstrated that only the simulations with a model combining both spherical and cylindrical scatterers within birefringent host medium reproduced the experimental trends in optical properties of the dermal layer (linear retardance, linear dichroism, and anisotropic linear depolarization) with layer thickness. Conclusions: Our studies prove that Mueller polarimetry provides relevant information not only on a size of dominant scatterers (e.g., cell nuclei versus subwavelength organelles) but also on its shape (e.g., cells versus collagen fibers). The latter is directly related to the state of extracellular collagen matrix, which is often affected by early pathology. Hence, using polarimetric data can help to increase the accuracy of diagnosis.

Mueller transmission microscopy has been used for both theoretical and experimental studies of anisotropic scattering biological tissue. In our prior study, the linear dependence of retardance and quadratic dependence of depolarization on thickness was demonstrated for a dermal layer of skin model. During the primary analysis of polarimetric images of histological cuts both epidermal and dermal layers were delineated manually in order to calculate the spatially averaged values of retardance and depolarization parameters. Consequently, these average values contained the contribution of outliers (noise, not correctly identified pixels, etc.) which produces large standard deviation and biased mean values of the parameters mentioned above. For preventing the errors, the normalized maps of optical properties were calculated pixel-wise taking into account local optical density (e. i. logarithm of M11 element of Mueller matrix at each image pixel) to compensate varying tissue thickness across the cut area. Furthermore, the DBSCAN (Density-based spatial clustering of applications with noise) algorithm was applied for segmentation of microscopic images using the normalized values of retardance, depolarization, and intensity. From the results of image segmentation, we could discriminate the regions of dermal and epidermal layers in Muller microscopic images of skin cuts more accurately and obtain more reliable values of tissue’s optical properties.