Skip to main content
SpringerLink
Log in
Book cover

Genetic Toxicology pp 121–141Cite as

The In Vitro Micronucleus Assay

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 817))

Abstract

The in vitro micronucleus test detects genotoxic damage in interphase cells. The in vitro micronucleus test provides an alterative to the chromosome aberration test, and because the in vitro micronucleus test examines cells at interphase, the assessment of micronuclei can be scored faster, as the analysis of damage is thought to be less subjective and is more amenable to automation.

Micronuclei may be the result of aneugenic (whole chromosome) or clastogenic (chromosome breakage) damage. This chapter provides methods for mononucleate and binucleate micronucleus tests and the addition of centromeric labelling and a non-disjunction assay to investigate any potential aneugenic mode of action.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Evans, H.J., Neary, C.J. and Williamson, F.S. (1959) The relative biological efficiency of single doses of fast neutrons and gamma rays on Vicia faba roots and the effect of oxygen, Part II. Chromosome damage: the production of micronuclei. Int Jnl Rad Biol 3, 216–229.

    Google Scholar 

  2. Schmid, W. (1975) The micronucleus test. Mutation Res 31, 9–15.

    Article  PubMed  CAS  Google Scholar 

  3. Fenech, M. and Morley, A.A. (1985) Measurement of micronuclei in lymphocytes. Mutation Res 147, 29–36.

    Article  PubMed  CAS  Google Scholar 

  4. OECD Guideline for Testing of Chemicals. No. 473: In Vitro Mammalian Chromosome Aberration Test (1997) www.oecd.org/env/testguidelines

  5. OECD Guideline for Testing of Chemicals. No 487 In Vitro Mammalian Cell Micronucleus Test (2010) www.oecd.org/env/testguidelines

  6. Migliore, L., Bocciardi, R., Macri, C. and Lo Jacono, F. (1993) Cytogenetic damage induced in human lymphocytes by four vanadium compounds and micronucleus analysis by fluorescence in situ hybridization with a centromeric probe. Mutation Res 319, 205–213.

    Article  PubMed  CAS  Google Scholar 

  7. Marshall, R.R., Murphy, M., Kirkland, D.J. and Bentley, K.S. (1996) Fluorescence in situ hybridisation (FISH) with chromosome-specific centromeric probes: a sensitive method to detect aneuploidy. Mutation Res 372, 233–245.

    Article  PubMed  CAS  Google Scholar 

  8. Zijno, P., Leopardi, F., Marcon, R. and Crebelli, R. (1996) Analysis of chromosome segregation by means of fluorescence in situ hybridization: application to cytokinesis-blocked human lymphocytes. Mutation Res 372 211–219.

    Article  PubMed  CAS  Google Scholar 

  9. Doherty, A.T., Ellard, S., Parry, E.M. and Parry, J.M. (1996) A study of aneugenic activity of trichlorofon detected by centromere-specific probes in human lymphoblastoid cell lines. Mutation Res 372, 221–231.

    Article  PubMed  CAS  Google Scholar 

  10. Ong, T., Mukhtar, M., Wolf, C.R. and Zeiger, E. (1980) Differential effects of cytochrome P450-inducers on promutagen activation capabilities and enzymatic activities of S-9 from rat liver. J Environ Pathol Toxicol 4, 55–65

    PubMed  CAS  Google Scholar 

  11. Elliott, B.M., Combes, R.D., Elcombe, C.R., Gatehouse, D.G., Gibson, G.G., Mackay, J.M. and Wolf, R.C. (1992) Alternatives to Aroclor 1254-induced S9 in in-vitro genotoxicity assays. Mutagenesis 7, 175–177.

    Article  PubMed  CAS  Google Scholar 

  12. Fellows, M.D., O’Donovan, M., Lorge, E. and Kirkland, D. (2008) Comparison of different methods for an accurate assessment of cytotoxicity in the in vitro micronucleus test. II. Practical Aspects with Toxic Agents. Mutation Res 655, 4–21.

    CAS  Google Scholar 

  13. Lorge, E., Hayashi, M., Albertini, S. and Kirkland, D. (2008) Comparison of different methods for an accurate assessment of cytotoxicity in the in vitro micronucleus test. Theoretical Aspects. Mutation Res 655, 1–3.

    Article  CAS  Google Scholar 

  14. Hozier, J., Sawyer, J., Moore, M., Howard, B. and Clive, D. (1998) Cytogenetic analysis of the L5178Y/TK+/− mouse lymphoma mutagenesis assay system. Mutation Res 84, 169–181

    Google Scholar 

  15. Varga, D., Johannes, T., Jainta, S., Suchuster, S., Schwarz-Boeger, U., Kiechle, M., Garcia, B.P. and Vogel, W. (2004) An automated scoring procedure for the micronucleus test. Mutagenesis 19, 391–397.

    Article  PubMed  CAS  Google Scholar 

  16. Countryman, P.I. and Heddle, J.A. (1976) The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes. Mutation Res 41, 321–332.

    Article  PubMed  CAS  Google Scholar 

  17. Schunck, C., Johannes, T., Varga, D., Lörch, T. and Plesch, A. (2004) New developments in automated cytogenetic imaging: unattended scoring of dicentric chromosomes, micronuclei, single cell gel electrophoresis, and fluorescence signals. Cytogenet Genome Res 104, 383–389.

    Article  PubMed  CAS  Google Scholar 

  18. Nüsse, M. and Kramer, J. (1984) Flow cytometric analysis of micronuclei found in cells after irradiation. Cytometry 5, 20–25.

    Article  PubMed  Google Scholar 

  19. Roman, D., Locher, F., Suter, W., Cordier, A., Bobadilla, M. (1998) Evaluation of a New Procedure for the Flow Cytometric Analysis of In Vitro, Chemically Induced Micronuclei in V79 Cells. Environmental and Molecular Mutagenesis 32, 387–396.

    Article  PubMed  CAS  Google Scholar 

  20. Lorge, E., Thybaud, V., Aardema, M.J., Oliver, J., Wakata, A., Lorenzon, G., Marzin, D. (2006) SFTG International collaborative Study on in vitro micronucleus test. I. General conditions of the study. Mutation Res 607, 13–36.

    CAS  Google Scholar 

  21. Clare, G., Lorenzon, G., Akhurst, L.C., Marzin, D., van Delft, J., Montero, R., Botta, A., Bertens, A., Cinelli, S., Thybaud, V. and Lorge, E. (2006) SFTG International collaborative study on the in vitro micronucleus test. II. Using human lymphocytes. Mutation Res 607, 37–60.

    CAS  Google Scholar 

  22. Miller, B., Potter-Locher, F., Seelbach, A., Stopper, H., Utesch, D. and Madle, S. (1998) Evaluation of the in vitro micronucleus test as an alternative to the in vitro chromosomal aberration assay: position of the GUM Working Group on the in vitro micronucleus test. Gesellschaft fur Umwelt-Mutations-forschung. Mutation Res 410, 81–116.

    CAS  Google Scholar 

  23. Bonassi, S., Fenech, M., Lando, C., Lin, Y.P., Ceppi, M., Chang, W.P., Holland, N., Kirsch-Volders, M., Zeiger, E., Ban, S., Barale, R., Bigatti, M.P., Bolognesi, C., Jia, C., Di Giorgio, M., Ferguson, L.R., Fucic, A., Lima, O.G., Hrelia, P., Krishnaja, A.P., Lee, T.K., Migliore, L., Mikhalevich, L., Mirkova, E., Mosesso, P., Muller, W.U., Odagiri, Y., Scarffi, M.R., Szabova, E., Vorobtsova, I., Vral, A. and Zijno, A. (2001) Human MicroNucleus Project: international database comparison for results with the cytokinesis-block micronucleus assay in human lymphocytes. I. Effect of laboratory protocol, scoring criteria and host factors on the frequency of micronuclei. Environ Mol Mutagen 37, 31–45.

    Article  PubMed  CAS  Google Scholar 

  24. Ellard, S. and Parry, E.M. (1993) A modified protocol for the cytochalasin B in vitro micronucleus assay using whole human blood or separated lymphocyte cultures. Mutagenesis 4, 317–320.

    Article  Google Scholar 

  25. Fenech, M. (2007) Cytokinesis-block micronucleus cytome assay. Nature Protocols 2, 1084–1104.

    Article  PubMed  CAS  Google Scholar 

  26. Fenech, M. (2000) The in vitro micronucleus technique. Mutation Res 455, 81–95.

    Article  PubMed  CAS  Google Scholar 

  27. Hoffman, W.P., Garriott, M.L. and Lee, C. (2003) In vitro micronucleus test, in Encyclopedia of Biopharmaceutical Statistics, (Chow, S., ed.), Marcel Dekker, Inc. New York, NY, pp. 463–467.

    Google Scholar 

  28. Garriott, M.L., Phelps, J.B. and Hoffman, W.P. (2002) A protocol for the in vitro micronucleus test I. Contributions to the development of a protocol suitable for regulatory submissions from an examination of 16 chemicals with different mechanisms of action and different levels of activity. Mutation Res 517, 123–134.

    Article  PubMed  CAS  Google Scholar 

  29. Kirsch-Volders, M., Sofuni, T., Aardema M., Albertini, S., Eastmond, D., Fenech, M., Ishidate Jr., M., Kirchner, S., Lorge, E., Morita, T., Norppa, H., Surrallés, J., Vanhauwaert, A. and Wakata, A., (2003) Report of the in vitro micronucleus assay working group. Mutat. Res. 540, 153–163

    CAS  Google Scholar 

Download references

Acknowledgements

The author would like to thank Julie Hayes and Mick Fellows for the helpful discussions and Karolina Chocian, Vicky Howarth, and Katie Clare for the technical assistance.

Author information

Authors and Affiliations

  1. Safety Assessment, AstraZeneca, Cheshire, UK

    Ann T. Doherty

Authors
  1. Ann T. Doherty
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ann T. Doherty .

Editor information

Editors and Affiliations

  1. Cedarmount 7, Lyndhurst, SO43 7ED, United Kingdom

    James M. Parry

  2. Cedarmount 7, Lyndhurst, SO43 7ED, Hampshire, United Kingdom

    Elizabeth M. Parry

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Doherty, A.T. (2012). The In Vitro Micronucleus Assay. In: Parry, J., Parry, E. (eds) Genetic Toxicology. Methods in Molecular Biology, vol 817. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-421-6_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-421-6_7

  • Published:

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-61779-420-9

  • Online ISBN: 978-1-61779-421-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation