Research Papers

Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy

[+] Author Affiliations
Jessica C. Mansfield, C. Peter Winlove, Julian Moger

University of Exeter, School of Physics, Stocker Road, Exeter, EX4 4QL, United Kingdom

Steve J. Matcher

University of Sheffield, The Kroto Institute, Broad Lane, Sheffield, S3 7HQ, United Kingdom

J. Biomed. Opt. 13(4), 044020 (July 15, 2008). doi:10.1117/1.2950318
History: Received September 13, 2007; Revised December 12, 2007; Accepted February 02, 2008; Published July 15, 2008
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Second harmonic generation (SHG) and two-photon fluorescence (TPF) microscopy is used to image the intercellular and pericellular matrix in normal and degenerate equine articular cartilage. The polarization sensitivity of SHG can be used directly to determine fiber orientation in the superficial 10to20μm of tissue, and images of the ratio of intensities taken with two orthogonal polarization states reveal small scale variations in the collagen fiber organization that have not previously been reported. The signal from greater depths is influenced by the birefringence and biattenuance of the overlying tissue. An assessment of these effects is developed, based on the analysis of changes in TPF polarization with depth, and the approach is validated in tendon where composition is independent of depth. The analysis places an upper bound on the biattenuance of tendon of 2.65×104. Normal cartilage reveals a consistent pattern of variation in fibril orientation with depth. In lesions, the pattern is severely disrupted and there are changes in the pericellular matrix, even at the periphery where the tissue appears microscopically normal. Quantification of polarization sensitivity changes with depth in cartilage will require detailed numerical models, but in the meantime, multiphoton microscopy provides sensitive indications of matrix changes in cartilage degeneration.

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© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Jessica C. Mansfield ; C. Peter Winlove ; Julian Moger and Steve J. Matcher
"Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy", J. Biomed. Opt. 13(4), 044020 (July 15, 2008). ; http://dx.doi.org/10.1117/1.2950318


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