Although X-ray microscopic computed tomography is widely used to assess the structural properties of polymeric tissue scaffolds its validity is dependent on the quality of the images obtained. Here, the role of resolution, integration time, image averaging, and X-ray power on the accurate determination of scaffold porosity, while aiming to minimize imaging time, was investigated. This work identified key parameters for optimization and a methodology to vary them to improve results. Based on this, guidelines were developed to assist in the selection of image acquisition parameters to allow rapid and accurate scaffold imaging as required for mass manufacture. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 100B: 360–367, 2012.