Imaging articular cartilage using second harmonic generation microscopy

Jessica C. Mansfield; C. Peter Winlove; Karen Knapp; Stephen J. Matcher

doi:10.1117/12.643830

23 February 2006 Imaging articular cartilage using second harmonic generation microscopy

Jessica C. Mansfield, C. Peter Winlove, Karen Knapp, Stephen J. Matcher

Proceedings Volume 6089, Multiphoton Microscopy in the Biomedical Sciences VI; 60891O (2006) https://doi.org/10.1117/12.643830
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

Sub cellular resolution images of equine articular cartilage have been obtained using both second harmonic generation microscopy (SHGM) and two-photon fluorescence microscopy (TPFM). The SHGM images clearly map the distribution of the collagen II fibers within the extracellular matrix while the TPFM images show the distribution of endogenous two-photon fluorophores in both the cells and the extracellular matrix, highlighting especially the pericellular matrix and bright 2-3μm diameter features within the cells. To investigate the source of TPF in the extracellular matrix experiments have been carried out to see if it may originate from the proteoglycans. Pure solutions of the following proteoglycans hyaluronan, chondroitin sulfate and aggrecan have been imaged, only the aggrecan produced any TPF and here the intensity was not great enough to account for the TPF in the extracellular matrix. Also cartilage samples were subjected to a process to remove proteoglycans and cellular components. After this process the TPF from the samples had decreased by a factor of two, with respect to the SHG intensity.

Citation Download Citation

Jessica C. Mansfield, C. Peter Winlove, Karen Knapp, and Stephen J. Matcher "Imaging articular cartilage using second harmonic generation microscopy", Proc. SPIE 6089, Multiphoton Microscopy in the Biomedical Sciences VI, 60891O (23 February 2006); https://doi.org/10.1117/12.643830

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