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Concentration dependence of the optical clearing effect created in muscle immersed in glycerol and ethylene glycol

We intended to characterize and compare the dependence between the concentration of two optical clearing agents and the effects created by them in muscle from rat. Using Ethylene glycol and Glycerol in three distinct concentrations, we expected to measure time evolution of the optical transmittance and variations created in tissue samples regarding mass, pH, thickness and histological parameters. Measuring natural state properties of tissue, we establish reference parameters to quantify variations in samples due to osmotic immersion treatment. Such variations were correlated with the optical clearing effect created in tissue and identified with time evolution of sample transmission. We observed for all the samples and agents studied that tissue transmission rises in time during the treatment with the osmotic solutions. Also, tissue thickness and refractive index show an increase, while the sample's pH lowers due to water loss inside tissue samples. Muscle fibres become more spatially separated after treatment due to osmotic impregnation inside the interstitial space. The variations described are stronger as the solution's concentration becomes higher. By comparing between results obtained with solutions of Ethylene glycol and Glycerol in the same concentration, we could verify similar effects but stronger when the Glycerol solution was applied.