In this study we showed that second-harmonic generation (SHG) microscopy combined with precise methods for images evaluation can be used to detect structural changes in the human ovarian stroma. Using a set of scoring (alignment collagen fibers, anisotropy, and correlation), found significant differences distribution organization fibers stroma component serous, mucinous, endometrioid mixed tumors as compared normal ovary tissue. This methodology was capable differentiate between cancerous...

Fluorophores are used to detect molecular expression by highly specific antigen-antibody reactions in fluorescence microscopy techniques. A portion of the fluorophore emits when irradiated with electromagnetic waves particular wavelengths, enabling its detection. Photobleaching irreversibly destroys fluorophores stimulated radiation within excitation spectrum, thus eliminating potentially useful information. Since this process may not be completely prevented, techniques have been developed...

Background 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 (THG) and lifetime imaging (FLIM) can detect morphological metabolic changes associated with cancer progression. Methodology/Principal Findings We obtained strong TPEF + SHG THG signals from fixed...

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. show that strong TPEF + SHG THG signals can be in fixed samples stained with hematoxylin eosin (H&E) stored for very long time, H&E staining enhanced the signal. then TPEF-SHG-THG microscopies file cancer obtain complementary information about...

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 metabolic changes associated with stroma epithelial transformation during the progression of cancer osteogenesis imperfecta (OI) disease. NLO microscopes provide complementary information about tissue microstructure, showing...

Endothelin (ET)-2, an ET family peptide, is highly expressed in intestine. However, the specific distribution and function of ET-2 remain unknown. We elucidated expression profile localization mouse gastrointestinal tract. Real-time PCR analysis revealed that gene tract healthy animals was relatively high colon. Immunohistochemical ET-2-like immunoreactivity mainly epithelial cells mucosa throughout intestinal animals. Intracellularly, concentrated close to basement membrane cells. A weak...

Abstract In this work, we proposed and built a multimodal optical setup that extends commercially available confocal microscope (Olympus VF300) to include nonlinear second harmonic generation (SHG) third (THG) (NLO) microscopy fluorescence lifetime imaging (FLIM). We explored all the flexibility offered by commercial capabilities. The allows image acquisition with confocal, brightfield, NLO/multiphoton FLIM imaging. Simultaneously, two‐photon excited (TPEF) SHG are well established in...

Background The confirmatory diagnosis of Osteogenesis Imperfecta (OI) requires invasive, commonly bone biopsy, time consuming and destructive methods. This paper proposes an alternative method using a combination two-photon excitation fluorescence (TPEF) second-harmonic generation (SHG) microscopies from easily obtained human skin biopsies. We show that this can distinguish subtypes OI. Methodology/Principal Findings Different aspects collagen microstructure fresh biopsies standard...

A vast number of human pathologic conditions are directly or indirectly related to tissular collagen structure remodeling. The nonlinear optical microscopy second-harmonic generation has become a powerful tool for imaging biological tissues with anisotropic hyperpolarized structures, such as collagen. During the past years, several quantification methods analyze and evaluate these images have been developed. However, automated semiautomated solutions necessary ensure objectivity...

Abstract Extracellular matrix (ECM) represents more than a mere intercellular cement. It is physiologically active in cell communication, adhesion and proliferation. Collagen the most abundant protein, making up to 90% of ECM, 30% total protein weight humans. Second-harmonic generation (SHG) microscopy an important tool study collagen organization ECM freshly unfixed tissues paraffin-embedded tissue samples. This manuscript aims review some applications SHG Oncologic Pathology, mainly...

Morphogenesis and architecture of a developing epithelium is controlled by both cell shape contacts, mediated spatially temporally regulated adhesion molecules. The authors study if E-cadherin functions as key factor epithelial epidermal in vivo. They apply whole-mount digital deconvolution microscopy to evaluate three-dimensional (3D) expression during skin morphogenesis Rhinella arenarum alteration model. Results show morphogenetic changes the 3D spatiotemporal pattern correlated with...

ABSTRACT Background: Colorectal cancer is one of the most prevalent pathologies worldwide whose prognosis linked to early detection. Colonoscopy gold standard for screening, and diagnosis usually made histologically from biopsies. Aiming reduce inspection diagnostic time as well biopsies resources involved, other techniques are being promoted conduct accurate in vivo colonoscopy assessments. Optical biopsy aims detect normal neoplastic tissues analysing autofluorescence spectrum based on...

Combined multimodal nonlinear optical (NLO) microscopies were used to detect and quantify morphological changes associated with stroma epithelial transformation in colon cancer. Our findings provide complementary information about tissue microstructure, displaying distinctive patterns between normal malignant human colon. Additionally, we have demonstrated the usefulness of using fixed tissues for disease diagnostic prognostic. The present work provides a framework NLO techniques as clinical...

We established a real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) system for the analysis of rat endothelin-1 (ET-1) and vasoactive intestinal contractor (VIC)/ET-2 gene expression. used this technique to examine expression levels in 16 different organs. ET-1 was observed all organs examined, while VIC mRNA detected some such as heart, lung, ovary, stomach, intestine. Ovary intestine express both at high levels, suggesting importance peptides these In addition,...

Colon cancer is one of the most diffused cancers in Western World, ranking third worldwide frequency incidence after lung and breast cancers. Even if it curable when detected treated early, a more accurate premature diagnosis would be suitable aim for both prognostic treatment. Combined multimodal nonlinear optical (NLO) microscopies, such as two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), harmonic (THG), lifetime imaging microscopy (FLIM) can used to detect...

Experimental models of pain in humans are crucial for understanding mechanisms. The most often used muscle involve the injection algesic substances, such as hypertonic saline solution or nerve growth factor induction delayed onset soreness (DOMS) by an unaccustomed exercise routine. However, these either invasive take substantial time to develop, and elicited level pain/soreness is difficult control. To overcome shortcomings, we propose elicit a localized application short-wave diathermy...

Murine induced colon cancer has been used to demonstrate that Second Harmonic Generation (SHG) microscopy images, combined with Two-Photon Excitation Fluorescence (TPEF) and specific quantization scoring methods allow distinguishing early alterations

To elucidate the physiological importance of endothelin-1 (ET-1) in mouse uterus, we investigated quantitative changes ET-1 mRNA levels uterus during estrous cycle, pregnancy and post-parturient period by use real-time PCR technique examined cellular distribution peptide immunohistochemical techniques. Low constant were observed from cyclic or pregnant mice. However, a significant increase was found immediately after parturition (day 0 postpartum) which then decreased gradually to basal...