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

Daniel S. Gareau, Paulo R. Bargo, William A. Horton, Steven Jacques

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

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 (M C) 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.

Original languageEnglish (US)
Pages (from-to)254-258
Number of pages5
JournalJournal of Biomedical Optics
Volume9
Issue number2
DOIs
StatePublished - Mar 2004

Fingerprint

cartilage
Fluorescence Spectrometry
Fluorescence spectroscopy
Cartilage
collagens
Green Fluorescent Proteins
Collagen
proteins
Proteins
Optical Fibers
fluorescence
Skin
Microspheres
spectroscopy
Optical fibers
Microscopes
optical fibers
Fluorescence
microscopes
pinholes

Keywords

  • Collagen
  • Confocal
  • EGFP
  • Fluorescence spectroscopy

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials
  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging
  • Radiological and Ultrasound Technology
  • Clinical Biochemistry

Cite this

Confocal fluorescence spectroscopy of subcutaneous cartilage expressing green fluorescent protein versus cutaneous collagen autofluorescence. / Gareau, Daniel S.; Bargo, Paulo R.; Horton, William A.; Jacques, Steven.

In: Journal of Biomedical Optics, Vol. 9, No. 2, 03.2004, p. 254-258.

Research output: Contribution to journalArticle

Gareau, Daniel S. ; Bargo, Paulo R. ; Horton, William A. ; Jacques, Steven. / Confocal fluorescence spectroscopy of subcutaneous cartilage expressing green fluorescent protein versus cutaneous collagen autofluorescence. In: Journal of Biomedical Optics. 2004 ; Vol. 9, No. 2. pp. 254-258.
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