The local concentration gradients of water and methanol within a Nafion (R) membrane are measured with a new microfluidic cell specially designed for depth measurement by confocal micro-Raman spectroscopy. The experimental concentration profiles of solvents, obtained in situ under cross-transport conditions, are fitted taking into account the contribution of the experimental set-up to the Raman response. This method allows to measure the concentration ratios of methanol and water at the solution-membrane interface as well as the real concentration gradients of these solvents within the membrane medium. These parameters are critical for a better understanding of membrane transport properties and cannot be directly measured by other techniques. Indeed, results here reported show that internal gradients differ from those that can be estimated from methods measuring external concentrations. (c) 2007 Elsevier B.V. All rights reserved.