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Journal of Biomedical Optics • Open Access

Elucidation of the mechanisms of optical clearing in collagen tissue with multiphoton imaging

Paper Abstract

Optical clearing (OC) is a promising method to overcome limitations in biomedical depth-resolved optical studies. Mechanisms of OC in purified bovine Achilles tendon, chicken skin, and chicken tendon were studied using time-lapsed, three-dimensional second harmonic generation (SHG) and two-photon fluorescence microscopic imaging. Quantified nonlinear optical measurements allowed temporal separation of two processes in collagen OC with glycerol. The first one is a fast process of tissue dehydration accompanied with collagen shrinkage and the second relatively slow process is glycerol penetration into the interfibrillar space of collagen alongside with CF swelling. The use of 50% glycerol induced less-expressed OC via partial substitution of water molecules with glycerol molecules. We also found that phosphate-buffered saline- and glycerol-treatments were reversible, and fiber morphology and SHG signal intensity were recovered after the removal of immersion agents. It was shown that tissue OC was a dynamic process and elucidation of its physical mechanisms may help choose optimal diagnostic, treatment, and modification regimes for collagen-based as well as other types of biomaterials.

Paper Details

Date Published: 2 April 2013
PDF: 9 pages
J. Biomed. Opt. 18(4) 046004 doi: 10.1117/1.JBO.18.4.046004
Published in: Journal of Biomedical Optics Volume 18, Issue 4
Show Author Affiliations
Vladimir A. Hovhannisyan, National Taiwan Univ. (Taiwan)
Po-Sheng Hu, National Taiwan Univ. (Taiwan)
Shean-Jen Chen, National Cheng Kung Univ. (Taiwan)
Chang-Seok Kim, Pusan National Univ. (Korea, Republic of)
Chen-Yuan Dong, National Taiwan Univ. (Taiwan)

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