The need for safe and non-invasive techniques for in situ structural and functional analysis of biological tissue has motivated widespread interest in optical spectroscopic-based imaging methods. Inclusion of spectroscopic information in the image data set can improve image contrast and, more importantly, provide for in situ histochemical tissue analysis via spectrochemical differences between tissue types. A promising approach utilizes the inherent spatial and spectral discrimination properties of the confocal Raman microscope (CRM) to isolate and extract 3D spectrochemical information from objects embedded in a scattering matrix. This study had several objectives. First, it is shown that a wax film immersed in a turbid solution of milk and water is a valid model system that replicates the elastic scattering properties and Raman activity typical of many biological tissues. Subsequently, the CRM spatial resolution (both axial and lateral) was measured as a function of solution turbidity with the tissue-equivalent phantom and the results were compared with those derived from a theoretical model of CRM image formation based on single scattering theory. The good agreement between the theoretical predictions and experimental results allowed limits to be established for confocal Raman spectroscopic imaging through a scattering tissue-like medium.
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Brenan et al. (1996) studied this question.