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Journal of Biomedical Optics

Single-wavelength reflected confocal and multiphoton microscopy for tissue imaging
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Paper Abstract

Both reflected confocal and multiphoton microscopy can have clinical diagnostic applications. The successful combination of both modalities in tissue imaging enables unique image contrast to be achieved, especially if a single laser excitation wavelength is used. We apply this approach for skin and corneal imaging using the 780-nm output of a femtosecond, titanium-sapphire laser. We find that the near-IR, reflected confocal (RC) signal is useful in characterizing refractive index varying boundaries in bovine cornea and porcine skin, while the multiphoton autofluorescence (MAF) and second-harmonic generation (SHG) intensities can be used to image cytoplasm and connective tissues (collagen), respectively. In addition, quantitative analysis shows that we are able to detect MAF from greater imaging depths than with the near-IR RC signal. Furthermore, by performing RC imaging at 488, 543, and 633 nm, we find that a longer wavelength leads to better image contrast for deeper imaging of the bovine cornea and porcine skin tissue. Finally, by varying power of the 780-nm source, we find that comparable RC image quality was achieved in the 2.7 to 10.7-mW range.

Paper Details

Date Published: 1 September 2009
PDF: 8 pages
J. Biomed. Opt. 14(5) 054026 doi: 10.1117/1.3247157
Published in: Journal of Biomedical Optics Volume 14, Issue 5
Show Author Affiliations
Wei-Liang Chen, National Taiwan Univ. (Taiwan)
Chen-Kuan Chou, National Taiwan Univ. (Taiwan)
Ming-Gu Lin, National Taiwan Univ. (Taiwan)
Yang-Fang Chen, National Taiwan Univ. (Taiwan)
Shiou-Hwa Jee, National Taiwan Univ. Hospital (Taiwan)
Hsian-Yuan Tan, Chang Gung Memorial Hospital (Taiwan)
Tsung-Hua Tsai, Far Eastern Memorial Hospital (Taiwan)
Ki Hean Kim, Pohang Univ. of Science and Technology (Korea, Republic of)
Daekeun Kim, Massachusetts Institute of Technology (United States)
Peter T. C. So, Massachusetts Institute of Technology (United States)
Sung-Jan Lin, National Taiwan Univ. Hospital (Taiwan)
Chen-Yuan Dong, National Taiwan Univ. (Taiwan)

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