Autofluorescence removal, multiplexing, and automated analysis methods for in-vivo fluorescence imaging

James R. Mansfield, Kirk W. Gossage, Clifford C. Hoyt, Richard M Levenson

Research output: Contribution to journalArticlepeer-review

244 Scopus citations


The ability to image and quantitate fluorescently labeled, markers in vivo has generally been limited by autofluorescence of the tissue. Skin, in particular, has a strong autofluorescence signal, particularly when excited in the blue or green wavelengths. Fluorescence labels with emission wavelengths in the near-infrared are more amenable to deep-tissue imaging, because both scattering and autofluorescence are reduced as wavelengths are increased, but even in these spectral regions, autofluorescence can still limit sensitivity. Multispectral imaging (MSI), however, can remove the signal degradation caused by autofluorescence while adding enhanced multiplexing capabilities. While the availability of spectral "libraries" makes multispectral analysis routine for well-characterized samples, new software tools have been developed that greatly simplify the application of MSI to novel specimens.

Original languageEnglish (US)
Article number041207
JournalJournal of Biomedical Optics
Issue number4
StatePublished - Jul 2005
Externally publishedYes


  • Autofluorescence removal
  • Fluorescence
  • In-vivo imaging
  • Quantum dots
  • Small animals
  • Spectral imaging

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


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