Research Papers

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

[+] Author Affiliations
Wanxin Sun

Institute of Bioengineering and Nanotechnology, The Nanos #4-01, 31 Biopolis Way, Singapore, Singapore 138669

Shi Chang

Central-South University, Xiangya Hospital, Department of General Surgery, Changsha, Hunan 410008, China and National University of Singapore, Department of Physiology, MD9, 2 Medical Drive, Singapore, Singapore 117597

Dean C. S. Tai

Institute of Bioengineering and Nanotechnology, The Nanos #4-01, 31 Biopolis Way, Singapore, Singapore 138669

Nancy Tan

National University of Singapore, Department of Physiology, MD9, 2 Medical Drive, Singapore, Singapore 117597 and K. K. Women’s and Children’s Hospital, Department of Paediatrics, 100 Bukit Timah Road, Singapore, Singapore 229899

Guangfa Xiao

Central-South University, Xiangya Hospital, Department of General Surgery, Changsha, Hunan 410008, China and National University of Singapore, Department of Physiology, MD9, 2 Medical Drive, Singapore, Singapore 117597

Huihuan Tang

Central-South University, Xiangya Hospital, Department of General Surgery, Changsha, Hunan 410008, China

Hanry Yu

Institute of Bioengineering and Nanotechnology, The Nanos #4-01, 31 Biopolis Way, Singapore, Singapore 138669 and

National University of Singapore, Department of Physiology and Graduate Programme in Bioengineering, Graduate School for Integrative Science and Engineering and Tissue-Engineering Programme, DSO Labs, Singapore, Singapore 117597 and Singapore-MIT Alliance, E4-04-10, 4 Engineering Drive 3, Singapore, Singapore 117576 and Massachusetts Institute of Technology, Department of Mechanical Engineering, Cambridge, Massachusetts 02139

J. Biomed. Opt. 13(6), 064010 (December 15, 2008). doi:10.1117/1.3041159
History: Received March 21, 2008; Revised July 18, 2008; Accepted August 28, 2008; Published December 15, 2008
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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.

Figures in this Article
© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Wanxin Sun ; Shi Chang ; Dean C. S. Tai ; Nancy Tan ; Guangfa Xiao, et al.
"Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies", J. Biomed. Opt. 13(6), 064010 (December 15, 2008). ; http://dx.doi.org/10.1117/1.3041159


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