Design of a confocal fluorescence spectrometer for transdermal observation of green fluorescent protein expression in cartilage

D. S. Gareau, H. Ma, W. A. Horton, S. L. Jacques

Research output: Contribution to journalConference article

Abstract

We have developed a noninvasive confocal fluorescence spectrometer to measure in vivo fluorescence spectra in mice. Our motivation is the study of the healing process of wounded cartilage in the murine model. The spectrometer measures the expression of green fluorescent protein (GFP) which is linked to the promoter of type II collagen as a marker for the expression of the chondrocyte phenotype. The confocal system uses an argon ion laser (488 nm) to excite fluorescence in a confocal volume about 200 um below the skin surface in the cartilage of the xyphoid process. In vitro studies showed how the refractive mismatch at the surface boundary and the light scattering of the tissue affect the depth and size of the confocal volume. A thin fluorescent layer was detected through a 130 um tissue phantom (murine skin) by moving the confocal volume axially through the phantom. The presence of skin increased the axial full width at half maximum of the confocal response from 7 um to 72 um. In conclusion, we achieve a low-spatial-resolution confocal spectrometer, which yields a fluorescence spectrum that maximizes the GFP fluorescence of cartilage and minimizes the skin autofluorescence.

Original languageEnglish (US)
Pages (from-to)132-135
Number of pages4
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume4261
DOIs
StatePublished - Jan 1 2001
EventThree-Dimensional and Multidimensional Microscopy: Image Asquisition and Processin VIII - San Jose, CA, United States
Duration: Jan 21 2001Jan 22 2001

Keywords

  • Axial point spread function
  • Confocal response
  • Confocal volume
  • Murine skin

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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