Development of the large scale digital cell analysis system

Quantitative analysis of large numbers of cells exposed to various stresses would be facilitated by an automated microscope system designed to monitor up to 1000 microscope fields over time intervals of up to one month. The Large Scale Digital Cell Analysis System (LSDCAS) was designed by the authors to provide such analysis capabilities. This report presents a description of the development of the LSDCAS and its application to the study of cells undergoing radiation-induced mitotic catastrophe. A detailed description of the data acquisition algorithms used and the hardware configuration underlying the system is presented. Quantitative analysis of colony formation of cells exposed to 5 Gy X irradiation is presented. Upon analysis, it was found that the plating efficiency in irradiated populations agreed with that obtained using conventional colony-formation assays, indicating the accuracy of LSDCAS in the determination of cell clonogenicity. An unexpected finding was that fully 50% of the surviving colonies exhibited mitotic catastrophe that persisted throughout colony formation. Ongoing extension of the capabilities of the LSDCAS using image segmentation techniques applied to a variety of cellular endpoints is then briefly described.