Abstract
BackgroundNonlinear 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.Methodology/Principal FindingsWe 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.Conclusions/SignificanceNLO 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.
Highlights
Epithelial ovarian cancers (EOC) constitute 90% of ovarian cancers and are insidious in nature because they are not detected and do not show signs and symptoms of the disease until late stages
The objective of this paper is to show the advantages of Non-Linear Optical (NLO) Microscopy to observe the carcinogenetic changes of EOCs
In this paper we show that Nonlinear optical (NLO) microscopy including Two-Photon Excited Fluorescence (TPEF), Second/Third Harmonic Generation (SHG/third-harmonic generation (THG)) and Fluorescence Lifetime Imaging (FLIM) microscopies of EOC fixed stained and unstained samples provide a set of complementary important information to develop indicators of the disease stage and progression
Summary
Epithelial ovarian cancers (EOC) constitute 90% of ovarian cancers and are insidious in nature because they are not detected and do not show signs and symptoms of the disease until late stages. This entity is responsible for the highest mortality among gynecologic cancers [1], with an estimated number of 21,990 new cases and 15,460 deaths in 2011 in the United States [2]. Serous adenocarcinomas often invade through the ovarian capsule and grow on the surface of the ovary This type of carcinoma is usually large, often bilateral, and is composed in part of papillae lined by stratified cells of serous type. 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
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