Angelo Pierangelo

Ex-vivo characterization of human colon cancer by Mueller polarimetric imaging

Cancerous and healthy human colon samples have been analyzed ex-vivo using a multispectral imaging Mueller polarimeter operated in the visible (from 500 to 700 nm) in a backscattering configuration with diffuse light illumination. Three samples of Liberkühn colon adenocarcinomas have been studied: common, mucinous and treated by radiochemotherapy. For each sample, several specific zones have been chosen, based on their visual staging and polarimetric responses, which have been correlated to the histology of the corresponding cuts. The most relevant polarimetric images are those quantifying the depolarization for incident linearly polarized light. The measured depolarization depends on several factors, namely the presence or absence of tumor, its exophytic (budding) or endophytic (penetrating) nature, its thickness (its degree of ulceration) and its level of penetration in deeper layers (submucosa, muscularis externa and serosa). The cellular density, the concentration of stroma, the presence or absence of mucus and the light penetration depth, which increases with wavelength, are also relevant parameters. Our data indicate that the tissues with the lowest and highest depolarizing powers are respectively mucus-free tumoral tissue with high cellular density and healthy serosa, while healthy submucosa, muscularis externa as well as mucinous tumor probably feature intermediate values. Moreover, the specimen coming from a patient treated successfully with radiochemotherapy exhibited a uniform polarimetric response typical of healthy tissue even in the initially pathological zone. These results demonstrate that multi-spectral Mueller imaging can provide useful contrasts to quickly stage human colon cancer ex-vivo and to distinguish between different histological variants of tumor.

Polarimetric imaging of uterine cervix: a case study

We present a preliminary investigation of macroscopic polarimetric imaging of uterine cervix. Orthogonal state contrast (OSC) images of healthy and anomalous cervices have been taken in vivo at 550 nm. Four ex vivo cervix samples have been studied in full Muller polarimetry, at 550 nm and 700 nm, and characterized in detail by standard pathology. One sample was totally healthy, another one carried CIN lesions at very early stage (CIN1) in its visible exocervical region, while for the other two samples more advanced (CIN3) lesions were present, together with visible glandular epithelium (ectropion). Significant birefringence has been observed in the healthy regions of all six samples, both in vivo and ex vivo. Standard treatments of the Mueller images of the ex vivo samples allowed to quantify both retardation and depolarization. Retardation reached 60° in healthy regions, and disappeared in the anomalous regions of the other three ex vivo samples. The depolarization power was largest in healthy regions, and lower in CINs and ectropion. Possible origins of the observed effects are briefly discussed.

Multispectral Mueller polarimetric imaging detecting residual cancer and cancer regression after neoadjuvant treatment for colorectal carcinomas

Abstract. This work is devoted to a first exploration of Mueller polarimetric imaging for the detection of residual cancer after neoadjuvant treatment for the rectum. Three samples of colorectal carcinomas treated by radiochemotherapy together with one untreated sample are analyzed ex vivo before fixation in formalin by using a multispectral Mueller polarimetric imaging system operated from 500 to 700 nm. The Mueller images, analyzed using the Lu-Chipmann decomposition, show negligible diattenuation and retardation. The nonirradiated rectum exhibits a variation of depolarization with cancer evolution stage. At all wavelengths on irradiated samples, the contrast between the footprint of the initial tumor and surrounding healthy tissue is found to be much smaller for complete tumor regression than when a residual tumor is present, even at volume fractions of the order of 5%. This high sensitivity is attributed to the modification of stromal collagen induced by the cancer. The depolarization contrast between treated cancer and healthy tissue is found to increase monotonously with the volume fraction of residual cancer in the red part of the spectrum. Polarimetric imaging is a promising technique for detecting short-time small residual cancers, which is valuable information for pathological diagnosis and patient management by clinicians.

Polarimetric Imaging for Cancer Diagnosis and Staging

A medical imaging technique that relies on light polarization could become a fast and accurate optical method for detecting cancer and determining the stage of the disease.

The origins of polarimetric image contrast between healthy and cancerous human colon tissue

Experimentally measured spectral Mueller matrix images of ex vivo human colon tissue revealed the contrast enhancement between healthy and cancerous zones of colon specimen compared to unpolarized intensity images. Cancer development starts with abnormal changes which being not yet visible macroscopically may alter the polarization of reflected light. We have shown with experiments and modeling that light scattering by small (sub wavelength) scatterers and light absorption (mainly due to blood hemoglobin) are the key factors for observed polarimetric image contrast. These findings can pave the way for the alternative optical technique for the monitoring and early detection of cancer.

Ex vivo photometric and polarimetric multilayer characterization of human healthy colon by multispectral Mueller imaging

Healthy human colon samples were analyzed ex vivo with a multispectral imaging Mueller polarimeter operating from 500 to 700 nm in a backscattering configuration with diffuse light illumination impinging on the innermost tissue layer, the mucosa. The intensity and polarimetric responses were taken on whole tissues first and after progressive exfoliation of the outer layers afterwards. Moreover, these measurements were carried out with two different substrates (one bright and the other dark) successively placed beneath each sample, allowing a reasonably accurate evaluation of the contributions to the overall backscattered light by the various layers. For the shorter investigated wavelengths (500 to 550 nm) the major contribution comes from mucosa and submucosa, while for the longer wavelengths (650 to 700 nm) muscular tissue and fat also contribute significantly. The depolarization has also been studied and is found to be stronger in the red part of the spectrum, mainly due to the highly depolarizing power of the muscular and fat layers.

Impact of model parameters on Monte Carlo simulations of backscattering Mueller matrix images of colon tissue

Polarimetric imaging is emerging as a viable technique for tumor detection and staging. As a preliminary step towards a thorough understanding of the observed contrasts, we present a set of numerical Monte Carlo simulations of the polarimetric response of multilayer structures representing colon samples in the backscattering geometry. In a first instance, a typical colon sample was modeled as one or two scattering “slabs” with monodisperse non absorbing scatterers representing the most superficial tissue layers (the mucosa and submucosa), above a totally depolarizing Lambertian lumping the contributions of the deeper layers (muscularis and pericolic tissue). The model parameters were the number of layers, their thicknesses and morphology, the sizes and concentrations of the scatterers, the optical index contrast between the scatterers and the surrounding medium, and the Lambertian albedo. With quite similar results for single and double layer structures, this model does not reproduce the experimentally observed stability of the relative magnitudes of the depolarizing powers for incident linear and circular polarizations. This issue was solved by considering bimodal populations including large and small scatterers in a single layer above the Lambertian, a result which shows the importance of taking into account the various types of scatterers (nuclei, collagen fibers and organelles) in the same model.

Advanced Mueller Ellipsometry Instrumentation and Data Analysis

The main object of this chapter is to give an overview the possibilities offered by instruments capable of measuring full Mueller matrices in the field of optical characterization. We have chosen to call these instruments Mueller ellipsometers in order to highlight their close relation with instruments traditionally used in ellipsometry. We want to make clear to the reader the place that Mueller ellipsometry takes with respect to standard ellipsometry by showing the similarities but also the differences among these techniques, both in instrumentation and data treatment. To do so the chapter starts by a review of the optical formalisms used in standard and Mueller ellipsometry respectively. In order to highlight the particularities and the advantages brought by Mueller ellipsometry, a special section is devoted to the algebraic properties of Mueller matrices and to the description of different ways to decompose them. Matrix decompositions are used to unveil the basic polarimetric properties of a the sample when a precise model is not available. Then follows a description of the most common optical configurations used to build standard ellipsometers. Special attention is paid to show what can and what cannot be measured with them. On the basis of this knowledge it is shown the interest of measuring whole Mueller matrices, in particular for samples characterized by complex anisotropy and/or depolarization. Among the numerous optical assemblies able to measure full Mueller matrices, most of them are laboratory prototypes, and only very few have been industrialized so far. Because an extensive and comparative review of all the Mueller ellipsometric instruments developed to date is clearly out of the scope of this chapter, we limit our description to four Mueller ellipsometers, two imaging and two spectroscopic systems that have been developed by us in the past years. The technical description of the Mueller ellipsometers is accompanied by some examples of applications which, without being exhaustive, are representative of the type of analyses performed in ellipsometry, and also illustrate the advantages that can be brought by modern Mueller ellipsometers to optical metrology, materials science and biomedicine.

Spatial evolution of depolarization in homogeneous turbid media within the differential Mueller matrix formalism.

We show, through visible-range Mueller polarimetry, as well as numerical simulations, that the depolarization in a homogeneous turbid medium consisting of submicron spherical particles follows a parabolic law with the path-length traveled by light through the medium. This result is in full agreement with the phenomenological theory of the fluctuating medium within the framework of the differential Mueller matrix formalism. We further found that the standard deviations of the fluctuating elementary polarization properties of the medium depend linearly on the concentration of particles. These findings are believed to be useful for the phenomenological interpretation of polarimetric experiments, with special emphasis on biomedical applications.

Ex vivo Mueller polarimetric imaging of the uterine cervix: a first statistical evaluation

Abstract. Early detection through screening plays a major role in reducing the impact of cervical cancer on patients. When detected before the invasive stage, precancerous lesions can be eliminated with very limited surgery. Polarimetric imaging is a potential alternative to the standard screening methods currently used. In a previous proof-of-concept study, significant contrasts have been found in polarimetric images acquired for healthy and precancerous regions of excised cervical tissue. To quantify the ability of the technique to differentiate between healthy and precancerous tissue, polarimetric images of seventeen cervical conization specimens (cone-shaped or cylindrical wedges from the uterine cervix) are compared with results from histopathological diagnoses, which is considered to be the “gold standard.” The sensitivity and specificity of the technique are calculated for images acquired at wavelengths of 450, 550, and 600 nm, aiming to differentiate between high-grade cervical intraepithelial neoplasia (CIN 2-3) and healthy squamous epithelium. To do so, a sliding threshold for the scalar retardance parameter was used for the sample zones, as labeled after histological diagnosis. An optimized value of ∼83% is achieved for both sensitivity and specificity for images acquired at 450 nm and for a threshold scalar retardance value of 10.6 deg. This study paves the way for an application of polarimetry in the clinic.