Probing bio-tissue films by optical internal reflectivity: modeling and measurements.

In this paper, we develop a detailed theoretical model for the optical reflectivity of a bio-tissue film confined between two flat interfaces based on the anomalous-diffraction approximation. We consider bio-tissue films consisting of a few layers of spheroidal cells surrounded by extracellular medium. We explore numerically the predictions of our model and compare them with simple effective medium theories, sometimes used as a first attempt to understand the optical properties of biological media. Then, we fit the model to experimental reflectivity-versus-angle-of-incidence curves of confined whole-blood films measured in an internal reflection configuration. Measurements were performed by confining a drop of fresh blood between a prism and a coverslip. Our experimental results show that it is possible to measure the coherent reflectance with small enough error to infer microstructural parameters with a good precision. The errors in measuring the coherent reflectance depend on the reflectivity magnitude. For instance, for a reflectivity of about 0.3 the error is below 2%, and the refractive indices of cells and surrounding medium can be obtained with a precision better than 1%. These results also indicate that the present model can readily be used to figure out the physical changes experienced at the microscale in bio-tissue films during a physicochemical process.