Application of quantitative second-harmonic generation microscopy to dynamic conditions

Mohammad M. Kabir, V. V. G. Krishna Inavalli, Tung-Yuen Lau, Kimani C. Toussaint
Biomed. Opt. Express. 2013-10-21; 4(11): 2546
DOI: 10.1364/boe.4.002546

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We present a quantitative second-harmonic generation (SHG) imaging technique that
quantifies the 2D spatial organization of collagen fiber samples under dynamic
conditions, as an image is acquired. The technique is demonstrated for both a
well-aligned tendon sample and a randomly aligned, sparsely distributed collagen
scaffold sample. For a fixed signal-to-noise ratio, we confirm the applicability
of this method for various window sizes (pixel areas) as well as with using a
gridded overlay map that allows for correlations of fiber orientations within a
given image. This work has direct impact to in vivo biological studies by
incorporating simultaneous SHG image acquisition and analysis.

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