Fluorescence-based imaging has an enormous impact on our understanding of biological systems. However, in vivo fluorescence imaging is greatly influenced by tissue scattering. A better understanding of this dependence can improve the potential of noninvasive in vivo fluorescence imaging. In this article, we present a diffusion model, based on an existing master-slave model, of isotropic point sources imbedded in a scattering slab, representing fluorophores within a tissue. The model was compared with Monte Carlo simulations and measurements of a fluorescent slide measured through tissue-like phantoms with different reduced scattering coefficients (0.5-2.5 mm À1 ) and thicknesses (0.5-5 mm). Results show a good correlation between our suggested theory, simulations and experiments; while the fluorescence intensity decays as the slab's scattering and thickness increase, the decay rate decreases as the reduced scattering coefficient increases in a counterintuitive manner, suggesting fewer fluorescence artifacts from deep within the tissue in highly scattering media.

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Diffuse reflectance technique is popular in the study of tissue physiology through the change in optical properties in a noninvasive manner. Diffuse reflected light intensity is commonly collected either from a single distance with spectral measurement or from a single wavelength with different spatial distances. Improving existing systems is necessary in order to obtain information from greater depths and in smaller volumes. In this paper, we propose a fast and compact fiber probe-based diffuse reflectance method for combining the spectral measurements in the range of 400–950 nm and spatial information up to 1.33 mm from the illumination source. First, we chose the most appropriate analysis model for the proposed distances between the fiber probe and tested it on solid phantoms with varying scattering and absorption components. The measurements are compared to the scattering coefficients according to Mie theory and the absorption according to spectrophotometer measurements. Next, we measured two-layer phantoms with constant scattering and absorption contrast in different layer thicknesses. We extracted the penetration depth from the measured effective absorption coefficient. In the near-IR, we were able to detect the absorption coefficient of the bottom phantom layer behind a top layer of up to 5 mm. We achieved a maximum penetration depth of 5.7 mm for 5 mm top layer thickness at 900 nm. Our fiber probe diffuse reflectance system can be used in the near future for skin lesion detection in clinical studies.

Tattoos are highly trendy in western culture, but many people regret their tattoos for many reasons. It is essential to be aware of the ink location in advance to reduce the long and short-term side effects. In this study, diffuse reflectance (DR) experiments were conducted on two-layer (2L) tissue-mimicking phantoms, where ink was sandwiched between the layers. An appreciable difference in the DR profile was found between the 2L phantom with and without the tattoo ink using the crossover point (Cp) method. Our technique was applied to ex vivo porcine skin. A point of intersection was found, between the skin and the tattooed skin. In the shorter wavelengths (500-600 nm), a distinguishable 2L behavior was found, and in longer wavelengths (600-850 nm), a single layer behavior was found between the tattooed skin before and after the intersection. In biological tissue, this Cp indeed finds the tattoo ink without harm to the surrounding skin.