A statistical approach for analyzing clonogenic survival data.

The assay of colony-forming efficiency is a mainstay in the measurement of cell response in vitro to many physical and chemical agents. Currently, data on colony-forming efficiency can be calculated in a variety of ways. Authors rarely describe in detail the methods used to determine the extent of biological variation within experiments. The use of standard methods of data analysis and presentation would improve interpretation of data and facilitate comparison between laboratories. Here we propose such a method. Binomial and Poisson probability theory were used to increase the accuracy of the estimate of the surviving fraction and to create an objective criterion for determining whether data obtained from serial dilutions of cell numbers used in the assay of colony-forming efficiency should be excluded or included for further analysis. The variability inherent in the calculation of surviving fraction was determined by using Fieller's theorem, a special statistical application for assessing ratios of estimates, to determine the 95% confidence interval. All calculations were done on a simple and commercially available spreadsheet program.