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An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry

An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry

Published on: May 13, 2019

Buccal micronucleus cytome assay.

Philip Thomas¹, Nina Holland, Claudia Bolognesi

¹CSIRO Human Nutrition, Adelaide, South Australia.

Nature Protocols

|May 16, 2009

Summary

The Buccal Micronucleus Cytome (BMCyt) assay offers a quick, minimally invasive method to analyze DNA damage and cell health in buccal tissue. This assay is valuable for epidemiological studies linking lifestyle and environmental factors to disease risk.

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Last Updated: Jun 23, 2026

An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry

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Published on: May 13, 2019

The Micronucleus Assay on Cryopreserved Whole Blood

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Automation of the Micronucleus Assay Using Imaging Flow Cytometry and Artificial Intelligence

Automation of the Micronucleus Assay Using Imaging Flow Cytometry and Artificial Intelligence

Published on: January 27, 2023

Area of Science:

Biomarker Discovery
Molecular Epidemiology
Genotoxicity Testing

Background:

The Buccal Micronucleus Cytome (BMCyt) assay is a key tool for assessing DNA damage and chromosomal instability in human buccal mucosal cells.
This assay provides insights into cell death and tissue regenerative potential, crucial for understanding disease etiology.
Biomarkers from the BMCyt assay are linked to increased risks of aging, cancer, and neurodegenerative conditions.

Purpose of the Study:

To detail a standardized protocol for the Buccal Micronucleus Cytome (BMCyt) assay.
To outline methods for buccal cell collection, slide preparation, and staining for microscopic analysis.
To provide clear scoring criteria for micronuclei and other nuclear anomalies.

Main Methods:

Buccal cell collection using a toothbrush, followed by single-cell suspension generation.
Slide preparation via cytocentrifugation, fixation, and Feulgen/Light Green staining.
Microscopic analysis (bright field and fluorescence) with detailed scoring for nuclear anomalies.

Main Results:

The protocol enables detailed analysis of DNA damage and chromosomal instability.
Established scoring criteria facilitate accurate assessment of micronuclei and other nuclear anomalies.
The entire process from cell collection to stained slide generation takes approximately 4 hours.

Conclusions:

The BMCyt assay is a robust and efficient method for evaluating DNA damage and cellular health in epidemiological studies.
This standardized protocol facilitates consistent and reliable assessment of genotoxicity and related health risks.
The assay's minimally invasive nature and relatively short duration make it suitable for large-scale human studies.