摘要
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(z_(f))versus increasing depth of focus zf behaved as R=ρexp(−μz_(f) 2G),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(μ_(s)[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 μ_(s) of dermis at about 300cm^(−1).DMSO and saline had only slight and inconsistent effects on g and μ_(s).
基金
the National Institutes of Health(R01-CA113947,R01-HL084013)in the USA
supported by the National Institutes of Health under the Ruth L.Kirschstein National Research Service Award 5-T32-CA106195-05,“Training in the Molecular Basis of Skin Pathobiology”,from the National Cancer Institute and the OHSU Cancer Institute
funded by the National Cancer Institute,CA-069533.
