Diffuse reflectance spectra are used to determine the optical properties of biological samples. In medicine and forensic science, the turbid objects under study often possess large absorption and/or scattering properties. However, data analysis is frequently based on the diffusion approximation to the radiative transfer equation, implying that it is limited to tissues where the reduced scattering coefficient dominates over the absorption coefficient. Nevertheless, up to absorption coefficients of 20 mm −1 at reduced scattering coefficients of 1 and 11.5 mm −1 , we observed excellent agreement (r 2 =0.994 ) between reflectance measurements of phantoms and the diffuse reflectance equation proposed by Zonios et al. [Appl. Opt.38, 6628–6637 (1999)], derived as an approximation to one of the diffusion dipole equations of Farrell et al. [Med. Phys.19, 879–888 (1992)]. However, two parameters were fitted to all phantom experiments, including strongly absorbing samples, implying that the reflectance equation differs from diffusion theory. Yet, the exact diffusion dipole approximation at high reduced scattering and absorption also showed agreement with the phantom measurements. The mathematical structure of the diffuse reflectance relation used, derived by Zonios et al. [Appl. Opt.38, 6628–6637 (1999)], explains this observation. In conclusion, diffuse reflectance relations derived as an approximation to the diffusion dipole theory of Farrell et al. can analyze reflectance ratios accurately, even for much larger absorption than reduced scattering coefficients. This allows calibration of fiber-probe set-ups so that the object’s diffuse reflectance can be related to its absorption even when large. These findings will greatly expand the application of diffuse reflection spectroscopy. In medicine, it may allow the use of blue/green wavelengths and measurements on whole blood, and in forensic science, it may allow inclusion of objects such as blood stains and cloth at crime scenes.
Current innovations in optical imaging, measurement techniques, and data analysis algorithms express the need for reliable testing and comparison methods. We present the design and characterization of silicone elastomer-based optical phantoms. Absorption is included by adding a green dye and scattering by adding TiO2 or SiO2 particles. Optical coherence tomography measurements demonstrate a linear dependence of the attenuation coefficient with scatterer concentration in the absence of absorbers. Optical transmission spectroscopy of the nonscattering absorbing phantoms shows a linear concentration dependent absorption coefficient. Both types of samples are stable over a period of 6 months. Confocal microscopy of the samples demonstrates a homogeneous distribution of the scatterers, albeit with some clustering. Based on layers with thicknesses as small as 50 µm, we make multifaceted structures resembling flow channels, (wavy) skin-like structures, and a layered and curved phantom resembling the human retina. Finally, we demonstrate the ability to incorporate gold nanoparticles within the phantoms. In conclusion, our phantoms are easy to make, are based on affordable materials, exhibit well-defined and controllable thickness, refractive index, absorption, and scattering coefficients, are homogeneous, and allow the incorporation of novel types of nanoparticle contrast agents. We believe our phantoms fulfill many of the requirements for an "ideal" tissue phantom, and will be particularly suited for novel optical coherence tomography applications.