While basic principles of microtubule organization are well understood, much remains to be learned about the extent and significance of variation in that organization among cell types and conditions. Large numbers of images of microtubule distributions for many cell types can be readily obtained by high throughput fluorescence microscopy but direct estimation of the parameters underlying the organization is problematic because it is difficult to resolve individual microtubules present at the microtubule-organizing center or at regions of high crossover. Previously, we developed an indirect, generative model-based approach that can estimate such spatial distribution parameters as the number and mean length of microtubules. In order to validate this approach, we have applied it to 3D images of NIH 3T3 cells expressing fluorescently-tagged tubulin in the presence and absence of the microtubule depolymerizing drug nocodazole. We describe here the first application of our inverse modeling approach to live cell images and demonstrate that it yields estimates consistent with expectations.