The microtubule network plays critical roles in many cellular processes, and quantitative models of how its organization varies across cell types and conditions are required for understanding those roles and as input to cell simulations. High-throughput image acquisition technologies are potentially valuable for this purpose, but do not provide sufficient resolution for current analysis methods that rely on tracing of individual microtubules. We describe a parametric conditional model of microtubule distribution that can generate a microtubule network in intact cells using a persistent random walk approach. The model parameters are physically meaningful as they directly describe the spatial distribution of microtubules and include the number of microtubules as well as the mean of the length distribution. We also present an indirect method for estimating the parameters of the model from three-dimensional fluorescence microscope images of cells that relies on comparing acquired images with simulated images generated from the model. Our results show that our method can reasonably recover parameters for a given query image, and we present the distributions of parameters estimated by our method for a collection of HeLa cell images. (c) 2010 International Society for Advancement of Cytometry.