Assessment of optical clearing agents using reflectance-mode confocal scanning laser microscopy

Ravikant Samatham, Kevin G. Phillips, Steven L. Jacques

Research output: Contribution to journalArticlepeer-review

39 Scopus citations


The mechanism of action of clearing agents to improve optical imaging of mouse skin during reflectance-mode confocal microscopy was tested. The dermal side of excised dorsal mouse skin was exposed for one hour to saline, glycerin, or 80% DMSO, then the clearing agent was removed and the dermis placed against a glass cover slip through which a confocal microscope measured reflectance at 488 nm wavelength. An untreated control was also measured. The axial attenuation of reflectance signal, R(zf) versus increasing depth of focus z f behaved as R = ρexp(-μzf2G), where ρ is tissue reflectivity and μ is attenuation [cm-1]. The factor 2G accounts for the in/out path of photons, and the numerical aperture of the lens. The ρ, μ data were mapped to values of scattering coefficient (μs [cm-1]) and anisotropy of scattering (g). Images showed that glycerin significantly increased the g of dermis from about 0.7 to about 0.99, with little change in the μs of dermis at about 300 cm-1. DMSO and saline had only slight and inconsistent effects on g and μs.

Original languageEnglish (US)
Pages (from-to)183-188
Number of pages6
JournalJournal of Innovative Optical Health Sciences
Issue number3
StatePublished - Jul 2010


  • Optical properties
  • anisotropy
  • skin

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Medicine (miscellaneous)
  • Atomic and Molecular Physics, and Optics
  • Biomedical Engineering


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