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Special Section on Optics of Human Skin

Confocal fluorescence spectroscopy of subcutaneous cartilage expressing green fluorescent protein versus cutaneous collagen autofluorescence

[+] Author Affiliations
Daniel S. Gareau, Paulo R. Bargo, William A. Horton, Steven L. Jacques

Oregon Health and Science University, Departments of Biomedical Engineering and Dermatology, Shriners Hospital for Crippled Children, Portland, Oregon?97239

J. Biomed. Opt. 9(2), 254-258 (Mar 01, 2004). doi:10.1117/1.1645798
History: Received Apr. 17, 2003; Revised Oct. 24, 2003; Accepted Oct. 24, 2003; Online March 22, 2004
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Optically monitoring the expression of green fluorescent protein (GFP) in the cartilage underlying the skin of a mouse allows tracking the expression of the chondrocyte phenotype. This paper considers how confocal microscopy with spectral detection can sense GFP fluorescence in the cartilage despite light scattering and collagen autofluorescence from the overlying skin. An in vivo experiment tested the abilities of a topical optical fiber measurement and a confocal microscope measurement to detect GFP in cartilage under the skin versus the collagen autofluorescence. An ex vivo experiment tested the ability of a confocal microscope without and with its pinhole to detect a fluorescent microsphere underneath an ex vivo skin layer versus the collagen autofluorescence. In both systems, spectroscopic detection followed by linear analysis allowed spectral discrimination of collagen autofluorescence (MC) and the subdermal green fluorescence (MG) due to either GFP or the microsphere. Contrast was defined as MG/(MG+MC). The in vivo contrast for GFP using optical fiber and confocal measurements was 0.16 and 0.92, respectively. The ex vivo contrast for a fluorescent microsphere using a confocal system without and with a pinhole was 0.13 and 0.48, respectively. The study demonstrates that a topical optical fiber measurement is affected by collagen autofluorescence, while a confocal microscope can detect subdermal fluorescence while rejecting collagen autofluorescence. © 2004 Society of Photo-Optical Instrumentation Engineers.

© 2004 Society of Photo-Optical Instrumentation Engineers

Citation

Daniel S. Gareau ; Paulo R. Bargo ; William A. Horton and Steven L. Jacques
"Confocal fluorescence spectroscopy of subcutaneous cartilage expressing green fluorescent protein versus cutaneous collagen autofluorescence", J. Biomed. Opt. 9(2), 254-258 (Mar 01, 2004). ; http://dx.doi.org/10.1117/1.1645798


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