Nonlinear optical microscopy of articular cartilage

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Presentation transcript:

Nonlinear optical microscopy of articular cartilage Alvin T. Yeh, Ph.D., Marie J. Hammer-Wilson, M.S., David C. Van Sickle, D.V.M., Ph.D., Hilary P. Benton, Ph.D., Aikaterini Zoumi, Ph.D., Bruce J. Tromberg, Ph.D., George M. Peavy, D.V.M., D.A.B.V.P. Osteoarthritis and Cartilage Volume 13, Issue 4, Pages 345-352 (April 2005) DOI: 10.1016/j.joca.2004.12.007 Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 1 Schematic of articular cartilage sample preparation for NLOM imaging. Images were obtained from articular and cut surface (asterisk). Osteoarthritis and Cartilage 2005 13, 345-352DOI: (10.1016/j.joca.2004.12.007) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 2 Histology of articular cartilage showing (A) full thickness, (B) superficial region and (C) boundary between non-calcified and calcified cartilage. Osteoarthritis and Cartilage 2005 13, 345-352DOI: (10.1016/j.joca.2004.12.007) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 3 Spectra of endogenous nonlinear optical signals in articular cartilage used for imaging. (A) Intensity image of chondrocytes within cartilage matrix. (B) Spectral filtering of SHG signal at 400nm specifically imaging collagen. (C) Spectral filtering of TPF signal at 520nm specifically imaging the chondron. Scale bar is 8μm. Osteoarthritis and Cartilage 2005 13, 345-352DOI: (10.1016/j.joca.2004.12.007) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 4 NLOM images (SHG and TPF) and SOFG stained histology of normal (A, B) and degenerative joint diseased (C) articular cartilage. Extracellular TPF corresponds with proteoglycan content (orange staining). Osteoarthritis and Cartilage 2005 13, 345-352DOI: (10.1016/j.joca.2004.12.007) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 5 Verhoeff's stained histology suggesting the presence of elastin (gray fibers) within the superficial layer. Depth-dependent intensity (SHG+TPF), TPF and SHG images showing ability to segment elastin-like fibers (TPF) and collagen (SHG) near the articular surface and matrix compositional and morphological changes with depth. Osteoarthritis and Cartilage 2005 13, 345-352DOI: (10.1016/j.joca.2004.12.007) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 6 NLOM images composed of (A) combined SHG and TPF, (B) SHG and (C) TPF signals obtained at 0.2μm intervals through a 32μm×30μm×14μm volume of the superficial zone of normal bovine articular cartilage are stacked to provide a 3-D image of the tissue. The images constructed here demonstrate that fluorescing fibrous structures of the superficial zone observed by TPF are distinct from collagen imaged by SHG. Osteoarthritis and Cartilage 2005 13, 345-352DOI: (10.1016/j.joca.2004.12.007) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 7 Histology and NLOM images of (A) normal, (B) early fibrillar matrical degeneration and (C) fibrocartilage of advanced degenerative joint disease. Intensity (column 1) and SHG NLOM images using vertically (column 2) and horizontally (column 3) polarized incident laser light are shown. Osteoarthritis and Cartilage 2005 13, 345-352DOI: (10.1016/j.joca.2004.12.007) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions