Quantitative analysis of biological tissues using Fourier transform-second-harmonic generation imaging

Raghu Ambekar Ramachandra Rao; Monal R. Mehta; Kimani C. Toussaint Jr.

doi:10.1117/12.841208

26 February 2010 Quantitative analysis of biological tissues using Fourier transform-second-harmonic generation imaging

Raghu Ambekar Ramachandra Rao, Monal R. Mehta, Kimani C. Toussaint Jr.

Proceedings Volume 7569, Multiphoton Microscopy in the Biomedical Sciences X; 75692G (2010) https://doi.org/10.1117/12.841208
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

We demonstrate the use of Fourier transform-second-harmonic generation (FT-SHG) imaging of collagen fibers as a means of performing quantitative analysis of obtained images of selected spatial regions in porcine trachea, ear, and cornea. Two quantitative markers, preferred orientation and maximum spatial frequency are proposed for differentiating structural information between various spatial regions of interest in the specimens. The ear shows consistent maximum spatial frequency and orientation as also observed in its real-space image. However, there are observable changes in the orientation and minimum feature size of fibers in the trachea indicating a more random organization. Finally, the analysis is applied to a 3D image stack of the cornea. It is shown that the standard deviation of the orientation is sensitive to the randomness in fiber orientation. Regions with variations in the maximum spatial frequency, but with relatively constant orientation, suggest that maximum spatial frequency is useful as an independent quantitative marker. We emphasize that FT-SHG is a simple, yet powerful, tool for extracting information from images that is not obvious in real space. This technique can be used as a quantitative biomarker to assess the structure of collagen fibers that may change due to damage from disease or physical injury.

Citation Download Citation

Raghu Ambekar Ramachandra Rao, Monal R. Mehta, and Kimani C. Toussaint Jr. "Quantitative analysis of biological tissues using Fourier transform-second-harmonic generation imaging", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 75692G (26 February 2010); https://doi.org/10.1117/12.841208

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