ColonyCountJ: A User-Friendly Image J Add-on Program for Quantification of Different Colony Parameters in Clonogenic Assay

Abstract

Objective: There are several commercial automatic colony counters available but they are relatively expensive and associated with several quantifications related problems. Some of the key problems in automated colony counting are clustering of colonies and edge effects. Thus the main objective of this study is to develop a user friendly program for determining colony parameters of clonogenic assay. Methods: In the present study, a watershed algorithm was employed along with other ImageJ tools to develop ImageJ based add-on program i.e ColonyCountJ to calculate various colony parameters. To test this program, we have used colonies obtained after exposure of MCF-7 cells to different doses of γ-radiation. Results: The results obtained using this program were compared with the manual counting as well as with automated counting provided by Oxford Optronix GelCount colony counter. It was found that our results corroborated well with the manual as well as with commercial program. As a whole, the performance of ColonyCountJ with regard to processing time and sensitivity was at par with the commercial colony counter. Conclusion: In conclusion, "ColonyCountJ" add-on program customized for ImageJ with optimized parameters is a reliable method for quantifying the colonies obtained from clonogenic assay. This program will be of great use for researchers working in the area of toxicology, radiation biology, and cancer biology.