Evaluation of a new procedure for the flow cytometric analysis of in vitro, chemically induced micronuclei in V79 cells.
Measurement of the frequency of micronuclei induced in cells by ionizing radiation or by chemical treatment is widely used to analyze cytogenetic damage. The microscopic scoring of micronuclei is a tedious and time-consuming procedure. Therefore, attempts have been made to automate micronuclei scoring by means of image analysis or flow cytometry. A new procedure for the flow cytometric analysis of chemically induced micronuclei in V79 Chinese hamster cells has been established in our laboratory. Debris was separated from micronuclei by means of a new gating procedure using area and width fluorescence of the stained suspension of micronuclei and nuclei. In order to test the sensitivity and specificity of this improved method of flow cytometric analysis, five well-known mutagenic compounds were tested. With the new technique, the frequency of micronuclei measured and analyzed corresponded well with results obtained by conventional microscopy. In addition, a large series of negative compounds, and weak, middle, and strong micronuclei inducers, were tested in order to establish criteria for discrimination between genotoxic and nongenotoxic compounds by flow cytometry. This new procedure for flow cytometric detection of micronuclei represents a quick, reliable, and relatively simple method for in vitro micronucleus testing.
Roman
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D
Preclinical Safety, Toxicology/Pathology, Novartis Pharma AG, Basel, Switzerland. danielle.roman@pharma.novartis.com
Locher
F
F
Suter
W
W
Cordier
A
A
Bobadilla
M
M
D000818:Animals; D001761:Bleomycin / Q000633:toxicity; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003520:Cyclophosphamide / Q000633:toxicity; D005020:Ethyl Methanesulfonate / Q000633:toxicity; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry / Q000379:methods*; D015162:Micronucleus Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D014747:Vinblastine / Q000633:toxicity; D014750:Vincristine / Q000633:toxicity
D009153:Mutagens; D001761:Bleomycin; D014750:Vincristine; D014747:Vinblastine; D003520:Cyclophosphamide; D005020:Ethyl Methanesulfonate