Experimental and simulated angular profiles of fluorescence and diffuse reflectance emission from turbid media

Given the wavelength dependence of sample optical properties and the selective sampling of surface emission angles by noncontact imaging systems, differences in angular profiles due to excitation angle and optical properties can distort relative emission intensities acquired at different wavelengths. To investigate this potentiality, angular profiles of diffuse reflectance and fluorescence emission from turbid media were evaluated experimentally and by Monte Carlo simulation for a range of incident excitation angles and sample optical properties. For emission collected within the limits of a semi-infinite excitation region, normalized angular emission profiles are symmetric, roughly Lambertian, and only weakly dependent on sample optical properties for fluorescence at all excitation angles and for diffuse reflectance at small excitation angles relative to the surface normal. Fluorescence and diffuse reflectance within the emission plane orthogonal to the oblique component of the excitation also possess this symmetric form. Diffuse reflectance within the incidence plane is biased away from the excitation source for large excitation angles. The degree of bias depends on the scattering anisotropy and albedo of the sample and results from the correlation between photon directions upon entrance and emission. Given the strong dependence of the diffuse reflectance angular emission profile shape on incident excitation angle and sample optical properties, excitation and collection geometry has the potential to induce distortions within diffuse reflectance spectra unrelated to tissue characteristics.