Colon adenocarcinoma diagnosis in human samples by multicontrast nonlinear optical microscopy of hematoxylin and eosin stained histological sections

Combined multimodal nonlinear optical (NLO) microscopies were used to detect and quantify morphological changes associated with stroma and epithelial transformation in colon cancer. Our findings provide complementary information about tissue microstructure, displaying distinctive patterns between normal and malignant human colon. Additionally, we have demonstrated the usefulness of using fixed tissues for the disease diagnostic and prognostic. The present work provides a framework for using NLO techniques as a clinical diagnostic tool for human colon cancer. NLO metrics could be applied to other disorders, which are characterized by abnormal cell proliferation and collagen assembly.Fil: Adur, Javier Fernando. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentina. National Institute of Science and Technology on Photonics Applied to Cell Biology; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Entre Ríos. Universidad Nacional de Entre Ríos. Centro de Investigaciones y Transferencia de Entre Ríos; ArgentinaFil: Bianchi, Mariana. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pelegati, Vitor B.. National Institute of Science and Technology on Photonics Applied to Cell Biology; BrasilFil: Viale, Silvia. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; ArgentinaFil: Izaguirre, María Fernanda. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; ArgentinaFil: Carvalho, Hernandes Faustino. Universidade Estadual de Campinas; BrasilFil: Cesar, Carlos L.. Universidade Estadual de Campinas; BrasilFil: Casco, Victor Hugo. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentin

Quantitative Changes In Human Epithelial Cancers And Osteogenesis Imperfecta Disease Detected Using Nonlinear Multicontrast Microscopy.

We show that combined multimodal nonlinear optical (NLO) microscopies, including two-photon excitation fluorescence, second-harmonic generation (SHG), third harmonic generation, and fluorescence lifetime imaging microscopy (FLIM) can be used to detect morphological and metabolic changes associated with stroma and epithelial transformation during the progression of cancer and osteogenesis imperfecta (OI) disease. NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for different types of human breast cancer, mucinous ovarian tumors, and skin dermis of patients with OI. Using a set of scoring methods (anisotropy, correlation, uniformity, entropy, and lifetime components), we found significant differences in the content, distribution and organization of collagen fibrils in the stroma of breast and ovary as well as in the dermis of skin. We suggest that our results provide a framework for using NLO techniques as a clinical diagnostic tool for human cancer and OI. We further suggest that the SHG and FLIM metrics described could be applied to other connective or epithelial tissue disorders that are characterized by abnormal cells proliferation and collagen assembly.17081407-

Recognition Of Serous Ovarian Tumors In Human Samples By Multimodal Nonlinear Optical Microscopy.

We used a multimodal nonlinear optics microscopy, specifically two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG∕THG) microscopies, to observe pathological conditions of ovarian tissues obtained from human samples. We show that strong TPEF + SHG + THG signals can be obtained in fixed samples stained with hematoxylin and eosin (H&E) stored for a very long time, and that H&E staining enhanced the THG signal. We then used the multimodal TPEF-SHG-THG microscopies in a stored file of H&E stained samples of human ovarian cancer to obtain complementary information about the epithelium∕stromal interface, such as the transformation of epithelium surface (THG) and the overall fibrillary tissue architecture (SHG). This multicontrast nonlinear optics microscopy is able to not only differentiate between cancerous and healthy tissue, but can also distinguish between normal, benign, borderline, and malignant specimens according to their collagen disposition and compression levels within the extracellular matrix. The dimensions of the layers of epithelia can also be measured precisely and automatically. Our data demonstrate that optical techniques can detect pathological changes associated with ovarian cancer.1609601

Six-wave mixing coherent anti-Stokes Raman scattering microscopy

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOAcquiring images of biological tissues and cells without the assistance of exogenous labels with a fast repetition rate and chemical specificity is what coherent anti-Stokes Raman Scattering (CARS) imaging offers. Nonresonant background (NRB) is one of the main drawbacks of the CARS microscopy technique because it limits the detection of weak Raman lines and the detection of low-concentration molecules. We show that a six-wave mixing process with two beams, which is a cascade effect of CARS, show better signal/NRB ratio and can be utilized for biological tissues imaging. The cascade CARS (CCARS) depends on chi-3 to the fourth power, instead of chi-3 squared as in the usual CARS signal||therefore, the contrast ratio with NRB is higher for CCARS than for CARS. We present analytic calculations showing that CCARS have better contrast over CARS in any situation. Comparison of the signals of both techniques generated on water-ethanol solutions confirm these results. Finally, we acquired CCARS images of fresh biological tissues, attesting that it is a useful tool for biological studies.9524072417FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO11/51959-005/51689-22013/16911-22014/50938-8312049/2014-5465699/2014-

Feminist Materialisms

We know how much matter and materiality influences, shapes and manipulates our becoming. Therefore how can we be anything but interested in feminist theories of materialism? We are interested in theorizing that opens a redefinition of how matter and materiality may be perceived. It is especially interesting when this theorizing also embraces the queerness in the world. This volume is a tribute to a new way of thinking about materiality, and represents a feminist voice in the ‘material turn’ that appears to be taking place in the social and human sciences

Optical Biomarkers Of Serous And Mucinous Human Ovarian Tumor Assessed With Nonlinear Optics Microscopies.

Nonlinear optical (NLO) microscopy techniques have potential to improve the early detection of epithelial ovarian cancer. In this study we showed that multimodal NLO microscopies, including two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), third-harmonic generation (THG) and fluorescence lifetime imaging microscopy (FLIM) can detect morphological and metabolic changes associated with ovarian cancer progression. We obtained strong TPEF + SHG + THG signals from fixed samples stained with Hematoxylin & Eosin (H&E) and robust FLIM signal from fixed unstained samples. Particularly, we imaged 34 ovarian biopsies from different patients (median age, 49 years) including 5 normal ovarian tissue, 18 serous tumors and 11 mucinous tumors with the multimodal NLO platform developed in our laboratory. We have been able to distinguish adenomas, borderline, and adenocarcinomas specimens. Using a complete set of scoring methods we found significant differences in the content, distribution and organization of collagen fibrils in the stroma as well as in the morphology and fluorescence lifetime from epithelial ovarian cells. NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for serous and mucinous ovarian tumors. The results provide a basis to interpret future NLO images of ovarian tissue and lay the foundation for future in vivo optical evaluation of premature ovarian lesions.7e4700

Experimental setup of a nonlinear optical microscopic system.

<p>CCD: charge-coupled device, AOM: acoustic-optic modulator, T₁: telescope, SP: short-pass filter, BP: band-pass filter, LP: long-pass filter, NDD: Non Descanned Detector. The SHG (red lines) and TPEF (green lines) are collected in a transmitted light configuration.</p

Best methods for quantitative analysis in fresh and fixed skin samples.

<p>Representative 3D maximum projection (40 images at intervals of 1 µm) of SHG images from fresh skin biopsies, A: normal skin, B: OI Type III (Patient E) and C: OI Type IV (Patient D). D: depth-dependent decay of the SHG signal analyzed to determine the collagen density in normal (black circle) and severe OI (Type III and IV -red circle). White asterisk indicate the selected regions where the plot was calculated as a function of depth. Each circle represent the mean value and standard deviation of three values corresponding to selected region. Texture analysis (E, F), using the gray-level co-occurrence matrix (GLCM). Energy values was calculated in dermis tissues versus distances pixels; ranging from 1 to 50 pixels (0.35 µm - 17.30 µm) in 0, 45, 90 and 135 deg directions of image (E: n = 12 normal, n = 3 mild OI, and n = 12 severe OI; and F: n = 12 normal, n = 3 OI Type I, n = 9 OI Type IV, n = 3 OI Type III). Pat: patients.</p

Optical Biomarkers of Serous and Mucinous Human Ovarian Tumor Assessed with Nonlinear Optics Microscopies

<div><h3>Background</h3><p>Nonlinear optical (NLO) microscopy techniques have potential to improve the early detection of epithelial ovarian cancer. In this study we showed that multimodal NLO microscopies, including two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), third-harmonic generation (THG) and fluorescence lifetime imaging microscopy (FLIM) can detect morphological and metabolic changes associated with ovarian cancer progression.</p> <h3>Methodology/Principal Findings</h3><p>We obtained strong TPEF + SHG + THG signals from fixed samples stained with Hematoxylin & Eosin (H&E) and robust FLIM signal from fixed unstained samples. Particularly, we imaged 34 ovarian biopsies from different patients (median age, 49 years) including 5 normal ovarian tissue, 18 serous tumors and 11 mucinous tumors with the multimodal NLO platform developed in our laboratory. We have been able to distinguish adenomas, borderline, and adenocarcinomas specimens. Using a complete set of scoring methods we found significant differences in the content, distribution and organization of collagen fibrils in the stroma as well as in the morphology and fluorescence lifetime from epithelial ovarian cells.</p> <h3>Conclusions/Significance</h3><p>NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for serous and mucinous ovarian tumors. The results provide a basis to interpret future NLO images of ovarian tissue and lay the foundation for future in vivo optical evaluation of premature ovarian lesions.</p> </div