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.
Biston
Marie-Claude
MC
Equipe d'Accueil no 2941 Rayonnement Synchrotron et Recherche Médicale', Unité IRM, CHU, BP 217, F-38043 Grenoble 09, France.
Corde
Stéphanie
S
Camus
Emmanuel
E
Marti-Battle
Ramon
R
Estève
François
F
Balosso
Jacques
J
D000465:Algorithms; D002452:Cell Count / Q000379:methods*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D045744:Cell Line, Tumor / Q000473:pathology / Q000528:radiation effects; D002470:Cell Survival / Q000502:physiology*; D003114:Colony-Forming Units Assay / Q000379:methods*; D003720:Densitometry / Q000379:methods*; D004307:Dose-Response Relationship, Radiation; D006258:Head and Neck Neoplasms / Q000473:pathology*; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods; D018307:Neoplasms, Squamous Cell / Q000473:pathology; D010363:Pattern Recognition, Automated; D014965:X-Rays