Abstract
Clonogenic assay is one of the most sensitive assays, widely used to evaluate the effects of antineoplastic agentsin vitro. A computer program was developed on an IBAS 2.0 Image Analysis System for automated quantiation of cell colonies and clone area on Petri dishes. The sensitivity of the clonogenic assay can be greatly increased by evaluating the mean area of the clones. The program gives an objective, accurate and fast evaluation of large samples. It is simple to use and offers a high degree of flexibility. Special algorithms and techniques have been implemented for good quantitation of both connected and well-separated colonies and to reduce the background noise and the general error rate. The principles and solutions presented are applicable to any other image analysis system.
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Abbreviations
- FBS:
-
fetal bovine serum
- ATCC:
-
American Type Culture Collection
- DDATHF:
-
5,10-dideazatetrahydrofolic acid
- PBS:
-
phosphate buffered saline
References
ATCC (1985) ATCC Quality Control Methods for Cell Line. First edition, RJ Hay (ed.), Rockville, MD, pp 12–15.
Buick RN, Pullano R & Trent JM (1985) Comparative properties of five human ovarian adenocarcinoma cell lines. Cancer Res 45: 3668–3676.
Erba E, Sen S, Sessa C, Vikhanskaya FL & D'Incalci M (1994) Mechanism of cytotoxicity of 5,10-dideazatetrahydrofolic acid in human ovarian carcinoma cellsin vitro and modulation of the drug activity by folic or folinic acid. Br. J. Cancer 69: 205–211.
Freshney RI (1987) Culture of Animal Cells. A Manual of Basic Technique. Alan R. Liss, New York, NY.
Gonzales RG & Woods RE (1992) Digital Image Processing. Addison-Wesley Publishing Company, Reading, MA.
Kelland LR, Mistry P, Abel G, Freidlos F, Loh SY, Roberts JJ & Harrap LR (1992) Establishment and characterization of anin vitro model of acquired resistance to cisplatin in a human testicular nonseminomatous germ cell line. Cancer Res. 52: 1710–1716.
Langdon SP, Lawrie SS, Hay FG, Hawkes MM, McDonald A, Hayward A, Schol DJ, Hilgers J, Leonard RCF & Smyth JF (1988) Characterization and properties of nine human ovarian adenocarcinoma cell lines. Cancer Res. 48: 6166–6172.
Parry RL, Chin TW & Donahoe PK (1991)Computer-aided cell colony counting. Biotechniques 10 (6): 772–774.
Pratt WK (1991) Digital Image Processing. Wiley Interscience, New York, NY.
Rupniak HT, Whelan RDH & Hill BT (1983) Concentration and time-dependent inter-relationships for antitumour drug cytotoxicities against tumour cellsin vitro. Int. J. Cancer 32: 7–12.
van den Berg-Bakker CAM, Hagemeijer A, Franken-Postma EM, Smit VTHBM, Kuppen PJK, van Ravenswaay Claasen HH, Cornelisse CJ & Schrier PI (1993) Establishment and characterization of 7 ovarian carcinoma cell lines and one granulosa tumor cell line: growth features and cytogenetics. Int. J. Cancer 53: 613–620.
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Bollea, A., Masseroli, M., Nicotra, A. et al. Automatic quantitation of cell colonies on petri dishes by computerized image analysis. Cytotechnology 17, 185–191 (1995). https://doi.org/10.1007/BF00749656
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DOI: https://doi.org/10.1007/BF00749656