5 November 2013 Quantifying the backscattering of second harmonic generation in tissues with confocal multiphoton microscopy
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The backward second harmonic generation (SHG) in mouse tissues is studied with a confocal multiphoton microscopy system. The total backward collected SHG (B-SHG) consists of the backward generated SHG and the backward-scattered forward-generated SHG (BS-SHG), which can be modeled by a Gaussian and a uniform distribution, respectively, at the confocal pinhole plane. By varying the pinhole size with a series of collection fibers, the proportion of the BS-SHG to the B-SHG and the proportion of BS-SHG to the forward generated SHG can be obtained. The approach is first validated by Monte Carlo simulation. It is then applied to two types of mouse tissues: mouse tail tendon and Achilles tendon. It is found that the BS-SHG contributes less to the B-SHG for the tail tendon than Achilles tendon with thicknesses of ∼300  μm . With the thickness of the Achilles tendon tissue increased to 1000 μm but the focal plane kept at the same depth, as high as ∼10% of the total forward SHG is backscattered and collected. The results indicate that BS-SHG may not be the major source of B-SHG in the tail tendon, but it may be the major source in the Achilles tendon. These methods and results provide a noninvasive method and supporting information for investigating the generation mechanism of SHG and help with optimizing backward SHG microscopy and spectroscopy measurements.
© 2013 Society of Photo-Optical Instrumentation Engineers (SPIE)
Mengzhe Shen, Yunxian Tian, Shau Poh Chong, Jianhua Zhao, Haishan Zeng, Shuo Tang, "Quantifying the backscattering of second harmonic generation in tissues with confocal multiphoton microscopy," Journal of Biomedical Optics 18(11), 115003 (5 November 2013). https://doi.org/10.1117/1.JBO.18.11.115003 . Submission:

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