Evaluation of Papanicolaou Stain for Studying Micronuclei in Buccal Cells Under Field Conditions

University of Maryland, Baltimore, Baltimore, Maryland, United States
Acta cytologica (Impact Factor: 1.56). 07/2006; 50(4):398-402. DOI: 10.1159/000325980
Source: PubMed


To compare Papanicolaou (Pap) and May-Grünwald Giemsa (MGG) stain as 2 techniques for staining for buccal mucosal cells to detect micronuclei (MN) infield studies.
Eighty cytologic smears (2 per individual) were taken from the buccal mucosa of 40 cigarette smokers recruited at a rural village in Egypt. Forty smears were stained with Pap stain and 40 with MGG stain. All were assessed for cellularity and scored for MN.
Pap stain was faster and easier to process and transport in the field study than was MGG stain. Regarding MGG smears, bacteria and cell debris masked the MN as compared to Pap smears, in which the fixative destroyed the bacteria and made the cell boundaries clearly demarcated. Using Pap stain, MN were seen easily in transparent cytoplasm.
Pap stain is the preferred method infield studies for scoring and detecting MN in cells of buccal mucosa.

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    • "According to the results of the detailed survey of the current methodological and data acquisition statuses of laboratories, that have published papers on the buccal MN assay (discussed at eighth workshop of the Human MicroNucleus (HUMN) project held at Antalya, Turkey, 2007), more than 50% of the laboratories which participated in the survey used Feulgen stain and only one of the laboratory had used H and E stain.[6] According to the same survey the second most common stain used was May-Grünwald-Giemsa (MGG) stain, however Ayyad et al.[15] found better results for counting of MNi using Pap stain as compared to MGG stain.Thus in the present study an attempt was made to compare Feulgen stain (the most commonly stain used for MN assay), Pap stain (the most commonly used stain in routine exfoliative cytology) and H and E stain (the most easily available stain). "
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    ABSTRACT: 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. 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). 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. 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. 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.
    Journal of Cytology 10/2012; 29(4):230-5. DOI:10.4103/0970-9371.103940 · 0.37 Impact Factor
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    ABSTRACT: The micronucleus (MN) assay in exfoliated buccal cells is a useful and minimally invasive method for monitoring genetic damage in humans. This overview has concluded that although MN assay in buccal cells has been used since the 1980s to demonstrate cytogenetic effects of environmental and occupational exposures, lifestyle factors, dietary deficiencies, and different diseases, important knowledge gaps remain about the characteristics of micronuclei and other nuclear abnormalities, the basic biology explaining the appearance of various cell types in buccal mucosa samples and effects of diverse staining procedures and scoring criteria in laboratories around the world. To address these uncertainties, the human micronucleus project (HUMN; see has initiated a new international validation project for the buccal cell MN assay similar to that previously performed using human lymphocytes. Future research should explore sources of variability in the assay (e.g. between laboratories and scorers, as well as inter- and intra-individual differences in subjects), and resolve key technical issues, such as the method of buccal cell staining, optimal criteria for classification of normal and degenerated cells and for scoring micronuclei and other abnormalities. The harmonization and standardization of the buccal MN assay will allow more reliable comparison of the data among human populations and laboratories, evaluation of the assay's performance, and consolidation of its world-wide use for biomonitoring of DNA damage.
    Mutation Research/Reviews in Mutation Research 05/2008; 659(1-2):93-108. DOI:10.1016/j.mrrev.2008.03.007 · 6.21 Impact Factor
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    ABSTRACT: Waterpipe smoking is popular in many parts of the world. Micronuclei (MN) evaluation in the exfoliated oral cells of smokers is a non-invasive technique for evaluation of possible tobacco harm. We aimed to assess whether MN levels are higher in waterpipe smokers than in never smokers. We examined oral smears of 128 adult male waterpipe smokers and 78 males who never smoked tobacco in rural Egypt. The total number of MN per 1000 cells per subject, and the number of MN-containing cells per individual were compared. We observed a higher level of total MN in waterpipe smokers (10 +/- 4) than in never smokers (4 +/- 2, p < 0.001). A similar difference was found for the mean number of affected cells per individual (8 +/- 3 vs. 4 +/- 1.62, p < 0.001). MN levels were not significantly dose related. This study is among the first to assess the association between waterpipe smoking and a cytogenetic measure of tobacco harm. The twofold increase in MN level is consistent with previous reports of MN in cigarette smokers. More research is needed to determine if such MN levels are predictive of future health consequences.
    Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 08/2008; 655(1-2):36-40. DOI:10.1016/j.mrgentox.2008.06.014 · 3.68 Impact Factor
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