Summary
The micronucleus test is a cytogenetic method for the screening of mutagens and carcinogens which exhibit clastogenic mechanisms of action. After application of clastogenic agents chromosomal fragments or even whole chromsomes can remain as conspicuous structures (micronuclei) in a small fraction of anucleated polychromatic erythrocytes which can be visually scored using a microscope following staining with May-Grünwald-Giemsa solutions. These time-consuming, painstaking, and tedious manual evaluations are often sources of unreliability and uncertainty. Here, a fluorescence technique is presented which applies DNA and protein fluorochromes to discriminate normal anucleated erythrocytes from micronucleated erythrocytes using a fluorescence microscope. This method is particularly tailored to be applied to flow cytometric instrumentation. Data obtained manually and automatically in flow show a strong linear correlation with high significance (r=0.96) as far as the percentage of micronucleated erythrocytes as an indicator for the mutagenicity of a given drug is concerned. These results have been obtained by means of the established clastogens cyclophosphamide and mitomycin C.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Boller K, Schmid W (1970) Chemische Mutagenesis beim Säuger. Das Knochenmark beim chinesischen Hamster als in vivo-Testsystem. Hämatologische Befunde nach Behandlung mit Trenimon. Hum Genet 11:35–54
Heddle JA (1973) A rapid in vivo test for chromosomal damage. Mutat Res 18:187–190
Hutter KJ, Stöhr M (1979) Two-parameter analysis of microbial cell constituents. Microbios Lett 10:121–128
King MT, Wild D (1979) Transplacental mutagenesis: The micronucleus test on fetal mouse blood. Hum Genet 51:183–194
Maier P, Schmid W (1976) Ten model mutagens evaluated by the micronucleus test. Mutat Res 40:325–338
Matter B, Schmid W (1971) Trenimon-induced chromosomal damage in bone-marrow cells of six mammalian species, evaluated by micronucleus test. Mutat Res 12:417–425
Melamed MR, Mullaney PF, Mendelsohn ML (1979) Flow cytometry and sorting. J Wiley, New York
Miller RC (1973) The micronucleus test as an in vivo cytogenetic method. Environ Health Perspect 6:167–170
Müller D, Langauer M, Rathenberg R, Strasser FF, Hess R (1972) Mikrokerntest sowie Chromosomenuntersuchungen an somatischen Zellen des chinesischen Hamsters nach Cyclophosphamidgabe. Verh Dtsch Ges Pathol 54:381–384
Salamone M, Heddle J, Stuart E, Katz M (1980) Towards an improved micronucleus test. Studies on 3 model agents, Mitomycin C, Cyclophosphamide and Dimethylbenzanthracene. Mutat Res 74:347–356
Schmid W (1973) Chemical mutagen testing on in vivo somatic mammalian cells. Agents Actions 3:77–85
Schmid W (1975) The micronucleus test. Mutat Res 31:9–15
Schmid W (1976) The micronucleus test for cytogenetic analysis. In: Hollaender A (ed) Chemical mutagens, principles and methods for their detection, Vol. 4. New York London, pp 31–53
Stöhr M, Vogt-Schaden M, Knobloch M, Vogel R, Futterman G (1978) Evaluation of eight fluorochrome combinations for simultaneous DNA-protein flow analysis. Stain Technol 53:205–215
Stöhr M, Goerttler Kl (1979) The Heidelberg flow analyzer and sorter (HEIFAS) approach on the prescreening of uterine cancer. J Histochem Cytochem 27:564–566
Wild D (1978) Cytogenetic effects in the mouse of 17 chemical mutagens and carcinogens evaluated by the micronucleus test. Mutat Res 56:319–327
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hutter, K.J., Stöhr, M. Rapid detection of mutagen induced micronucleated erythrocytes by flow cytometry. Histochemistry 75, 353–362 (1982). https://doi.org/10.1007/BF00496738
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00496738