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Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies

Author(s)
Sun, Wanxin; Chang, Shi; Tai, Dean C. S.; Tan, Nancy; Xiao, Guangfa; Tang, Huihuan; Yu, Hanry; ... Show more Show less
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Abstract
Liver fibrosis is associated with an abnormal increase in an extracellular matrix in chronic liver diseases. Quantitative characterization of fibrillar collagen in intact tissue is essential for both fibrosis studies and clinical applications. Commonly used methods, histological staining followed by either semiquantitative or computerized image analysis, have limited sensitivity, accuracy, and operator-dependent variations. The fibrillar collagen in sinusoids of normal livers could be observed through second-harmonic generation (SHG) microscopy. The two-photon excited fluorescence (TPEF) images, recorded simultaneously with SHG, clearly revealed the hepatocyte morphology. We have systematically optimized the parameters for the quantitative SHG/TPEF imaging of liver tissue and developed fully automated image analysis algorithms to extract the information of collagen changes and cell necrosis. Subtle changes in the distribution and amount of collagen and cell morphology are quantitatively characterized in SHG/TPEF images. By comparing to traditional staining, such as Masson’s trichrome and Sirius red, SHG/TPEF is a sensitive quantitative tool for automated collagen characterization in liver tissue. Our system allows for enhanced detection and quantification of sinusoidal collagen fibers in fibrosis research and clinical diagnostics.
Date issued
2008-12
URI
http://hdl.handle.net/1721.1/69125
Department
Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Mechanical Engineering
Journal
Journal of Biomedical Optics
Publisher
SPIE - International Society for Optical Engineering
Citation
Sun, Wanxin et al. “Nonlinear Optical Microscopy: Use of Second Harmonic Generation and Two-photon Microscopy for Automated Quantitative Liver Fibrosis Studies.” Journal of Biomedical Optics 13.6 (2008): 064010. Web. 16 Feb. 2012. © 2008 SPIE - International Society for Optical Engineering
Version: Final published version
ISSN
1083-3668
1560-2281

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