Analyzing MOA with In Vitro MultiFlow Assays

The In Vitro MultiFlow Assay is a rapid genetic toxicology screening tool. By assessing nuclear translocation of p53, the assay can differentiate between true genotoxicants and chemicals (or metabolites) that induce micronuclei due to cytotoxicity. Furthermore, measurement of γH2AX foci and mitotic perturbations informs on MOA, to help understand if a genotoxicant is operating via a non-DNA reactive mechanism.


Scientific Poster Utlizing High Content Assays

96-well plate with pipette used in na In vitro Multiflow assay

Identifying the Mode of Action of Potential Genotoxicants Using Nuclear Biomarkers

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Benefits of the in vitro MultiFlow assay

  • Screen libraries of compounds for potential genotoxic effects (using as little as 5 mg or 200 µL of a 100 mM solution).
  • Follow up a positive mammalian cell micronucleus assay test result to ensure the effect isn’t caused by cytotoxicity – eliminate misleading positives.
  • Build evidence for a non-DNA reactive MOA as needed.
  • Analyze four compounds concurrently in a series of 20 concentrations for high-throughput and lower testing costs.

Charles River scientists based in North America and Europe collaborate as a team and with clients to execute efficient genetic toxicology studies and IND-enabling programs that meet the requirements of regulatory agencies worldwide.

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Frequently Asked Questions (FAQs) for In Vitro MultiFlow® Assays

  • How does the MultiFlow® Assay help my genetic toxicology study?

    High content assays quickly screen limited amounts of test material for potential genotoxicity and predict the outcome of the GLP in vitro micronucleus test. If faced with a positive result from your in vitro micronucleus test result, the In Vitro MultiFlow assay can tell you if the micronuclei formed via an aneugenic mechanism. Hence, it is a high content FISH alternative

  • How predictive is MultiFlow to in vitro micronucleus outcomes?

    While there are exceptions to every rule, the In Vitro MultiFlow assay was designed to eliminate misleading positives in the TK6 micronucleus assay. Hence, some negative compounds in MultiFlow may be positive in the in vitro micronucleus assay, but the micronuclei likely formed due to cytotoxic, and not genotoxic, effects. Conversely, a positive result in MultiFlow is highly predictive of a positive outcome in the TK6 micronucleus assay, and there is growing evidence that the intensity of nuclear γH2AX directly correlates with micronucleus frequency.

  • If MultiFlow is negative, am I done testing?

    No. The In Vitro MultiFlow assay is another instrument in our genetic toxicity testing tool box and should be used according to the information provided herein. For regulatory purposes, the high content assay can inform on mode of action, but it does not replace the standard battery of OECD-compliant genetic toxicology assays.