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Updated: May 15, 2026
An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
Published on: May 13, 2019
A comparative study for selectivity of micronuclei in oral exfoliated epithelial cells
S Grover1, Abr Mujib, A Jahagirdar
1Department of Oral Pathology and Microbiology, Bapuji Dental College and Hospital, Davangere, Karnataka, India.
Background:
Micronucleus (MN) represents small, additional nuclei formed by the exclusion of chromosome fragments or whole chromosomes lagging at mitosis. MN rates, therefore, indirectly reflect chromosome breakage or impairment of the mitotic apparatus. During the last few decades, micronuclei ("MNi") in oral exfoliated epithelial cells are widely used as biomarkers of chromosomal damage, genome instability and cancer risk in humans. However, until now only little attention has been given to the effect of different staining procedures on the results of these MN assays.
Aim:
To compare the MNi frequencies in oral exfoliated epithelial cells using three different stains, i.e.,Feulgen stain, Papanicolaou stain (Pap) and hemotoxylin and eosin stain (H and E).
Materials And Methods:
Oral exfoliated cells from 45 cases of potentially malignant disorders (15 oral submucous fibrosis, 15 lichen planus and 15 leukoplakia) and 15 controls with healthy mucosa, were taken and MNi frequencies (No. of MNi/1000 cells) were compared using three different stains.
Results:
Mean MNi frequency in cases was found to be 3.8 with Feulgen stain, 16.8 with PAP and 25.9 with H and E. In controls, mean MNi frequency was 1.6 with Feulgen stain, 7.7 with PAP and 9.6 with H and E stain. Statistically significant value (P < 0.01) were observed when the three stains were compared together using Kruskal Walli's ANOVA test.
Conclusion:
Feulgen being a DNA-specific stain gave the least counts, although statistically significant results from the comparison of MNi frequency between cases and controls were obtained with all the three stains.
Insights
Staining methods significantly impact micronuclei (MNi) counts in oral cells, affecting biomarker reliability for chromosomal damage. Feulgen stain, a DNA-specific method, yielded lower MNi frequencies compared to Papanicolaou and H&E stains.
Area of Science:
- Oral pathology
- Cytogenetics
- Biomarker research
Background:
- Micronuclei (MNi) in oral exfoliated cells are established biomarkers for chromosomal damage, genome instability, and cancer risk.
- MN rates indirectly reflect chromosome breakage and mitotic apparatus function.
- The influence of different staining techniques on MN assay results remains underexplored.
Purpose of the Study:
- To evaluate and compare micronuclei frequencies in oral exfoliated epithelial cells using Feulgen, Papanicolaou (Pap), and Hematoxylin and Eosin (H&E) stains.
- To assess the impact of staining methodology on the detection of chromosomal damage biomarkers.
Main Methods:
- Oral exfoliated cells were collected from 45 patients with potentially malignant disorders (oral submucous fibrosis, lichen planus, leukoplakia) and 15 healthy controls.
- Micronuclei frequencies (MNi/1000 cells) were quantified using Feulgen, Pap, and H&E staining methods.
- Statistical analysis, including Kruskal-Wallis ANOVA, was employed to compare MNi frequencies across stains and groups.
Main Results:
- Mean MNi frequencies in patient cases were 3.8 (Feulgen), 16.8 (Pap), and 25.9 (H&E).
- Mean MNi frequencies in controls were 1.6 (Feulgen), 7.7 (Pap), and 9.6 (H&E).
- Statistically significant differences (P < 0.01) were observed in MNi counts among the three stains.
Conclusions:
- All three stains (Feulgen, Pap, H&E) detected statistically significant differences in MNi frequencies between cases and controls.
- Feulgen stain, being DNA-specific, provided the lowest MNi counts.
- Staining method choice can influence the quantitative assessment of micronuclei as biomarkers.
