Building quantitative stereology data files with scion image, a public domain image processing and analysis software

Two-dimensional data obtained from a histological cross-section of a tissue can be utilized to obtain three-dimensional information by the methods of quantitative stereology. The resulting quantitative information is useful in both experimental studies and whole-animal investigations for regulatory and safety purposes. Quantitative stereologic analysis requires considerable data collection and calculation and is thus practical only through the use of computer hardware and software. We have previously reported the development of a program, STEREO, which compiles data from carcinogenesis experiments, recording information from tissue sections for the estimation of the number of altered hepatic foci (AHF) per liver and the volume fraction of AHF in liver on a three-dimensional basis. The data file itself was built by measuring tissue and focal transections through a slide-reading process that involved the manual use of a digitizer. In order to increase the speed and efficiency of the analytical process, we have integrated the STEREO program with a public domain software, Scion Image. This software integration involves two portions: the building macros and the interface. Macros for quantitative stereology used in Scion Image were written to customize and simplify the measurement and to generate data needed for building each of the data files. An interface program, BuildFi.exe, was developed to receive data generated from Scion Image and to align sequential tissue plots from up to four serial sections stained with different markers. As a result, the user can store data on a disk in the format of the STEREO data files. By combining STEREO with Scion Image, the slide-reading process is simplified and can be performed automatically. It has proven to be more objective, time saving, and efficient than all earlier versions.