This paper demonstrates a technique that could prove useful for extracting three-dimensional (3D) models from a single two-dimensional (2D) digital in-line holographic microscopy (DIHM) recording. Multiple intensity images are reconstructed at a range of depths through a transmissive or partially transmissive scene recorded by DIHM. A two step segmentation of each of these reconstructed intensity images facilitates the construction of a data set of surfaces in 3D. First an adaptive thresholding step and then a border following step are implemented. The surfaces of segmented features are rendered in 3D by applying the marching cubes algorithm to polygonize the data set. Experimental results for a real world DIHM capture of a transmissive glass sample are presented to demonstrate this segmentation and visualization process. Â© 2009 SPIE.