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An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
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In Vitro Cytogenetic Assays: Chromosomal Aberrations and Micronucleus Tests.

Pasquale Mosesso1, Serena Cinelli2

  • 1Dipartimento di Scienze Ecologiche e Biologiche, Università degli Studi della Tuscia (Retired), Largo dell'Università, Viterbo, Italy. mosesso@unitus.it.

Methods in Molecular Biology (Clifton, N.J.)
|November 22, 2025
PubMed
Summary

This chapter details methods for detecting genotoxicity-induced chromosome damage in vitro. It covers structural aberrations at metaphase and the micronucleus test for interphase cells, crucial for environmental and clinical risk assessment.

Keywords:
Adherent cell lines (CHO, CHL, V79)Chromosome aberration assay protocolsChromosome aberrationsCytogenetic testsMicronucleus

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Area of Science:

  • Toxicology
  • Genetics
  • Molecular Biology

Background:

  • Chromosome damage is a key indicator of genetic damage.
  • Environmental and clinical studies rely on assessing genotoxicity.
  • Understanding genotoxic effects requires reliable detection methods.

Purpose of the Study:

  • To describe protocols for detecting chromosomal aberrations induced by genotoxic agents in vitro.
  • To present methods for analyzing both structural aberrations and micronuclei.
  • To outline criteria and statistical methods for positive result determination.

Main Methods:

  • Detailed protocols for in vitro chromosomal aberration analysis are provided.
  • The micronucleus test for detecting interphase chromosomal damage (fragments or whole chromosomes) is described.
  • Methods include assessment in the presence and absence of rat liver-derived metabolizing systems.

Main Results:

  • Structural chromosomal aberrations observable at metaphase are described.
  • The micronucleus test protocol for interphase cells is presented.
  • Criteria and statistical approaches for evaluating positive results are included.

Conclusions:

  • The chapter provides comprehensive methods for evaluating genotoxic effects on chromosomes.
  • Accurate detection of chromosomal damage is vital for risk assessment in various studies.
  • Standardized protocols ensure reliable identification of genotoxic agents.