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

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...

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Bibliographic Details
Published in:Journal of Biomedical Optics 2008-11, Vol.13 (6), p.064010-064017
Main Authors: Sun, Wanxin, Chang, Shi, Tai, Dean C. S, Tan, Nancy, Xiao, Guangfa, Tang, Huihuan, Yu, Hanry
Format: Article
Language:English
Subjects:
Algorithms
Animals
Artificial Intelligence
Automated
Collagens
fibrillar collagen
Fibrosis
Image Enhancement - methods
Image Interpretation, Computer-Assisted - methods
Liver
Liver Cirrhosis - pathology
liver fibrosis
Male
Microscopy
Microscopy, Fluorescence, Multiphoton - methods
Morphology
Nonlinear Dynamics
Pattern Recognition, Automated - methods
quantification
Rats
Rats, Wistar
Reproducibility of Results
second harmonic generation
Sensitivity and Specificity
Staining
Subtraction Technique
two-photon excited fluorescence
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Summary: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.
ISSN:1083-3668
1560-2281
DOI:10.1117/1.3041159