An objective method to measure cell survival by computer-assisted image processing of numeric images of Petri dishes

This work establishes an objective method to measure cell clonogenic survival by computer-assisted image processing using images of cell cultures fixed and stained in Petri dishes. The procedure, developed by Samba Technologies, consists of acquiring Petri dish pictures with a desktop scanner and analysing them by computer, using algorithms based on the 'top hat' filter. The results from the automated count for the cell line SQ20B are compared with those found by two observers, before and after normalization of the counting. After normalization, the shape of the survival curves of the 'manual' counting of the Petri dishes shows a good correlation between both observers. The software enables the small visible differences in count between observers to be eliminated. The comparison between the absolute number of colonies shows an increased difference between the two manual scorings that can be as great as 67 colonies, whereas the difference between the two automated counts is never greater than 8 colonies. These results demonstrate that the 'manual' count is inter- and intraobserver variable, whereas the automatic count performs reproducible cell colony counts, thereby minimizing user-generated bias. The large amount of data produced also gives information about cell and colony characteristics. Thus, this computer-assisted method has considerably improved the reliability of our statistical results.