A comparative study for qualitative and quantitative analysis of light and fluorescence microscopy stains in oral premalignant and malignant lesions

Abstract

Objectives: The present study highlights the use and benefit of fluorescence microscopy stains in easy diagnosis of oral premalignant and malignant lesions. This study was carried out to evaluate the efficiency of light microscopy using Papanicolaou stain (PAP) and May Grunwald Giemsa (MGG) stains and fluorescence microscopy using acridine orange (AO) and 4′,6-diamidino-2-phenyl indole (DAPI) stains. All smears have been qualitatively and quantitatively analyzed in premalignant and malignant lesions using cytology and cytomorphometry. Materials and Methods: The subjects included in this study were selected from patients suffering from premalignant and clinically suspicious malignant lesions. Specimens were collected from the most representative site in study groups and normal buccal mucosa in the control group. The smears were evaluated under fluorescence microscopy for AO and DAPI and under light microscopy for conventional PAP and MGG. We studied 5 HPF to compare the number of normal cells to dysplastic cells. These were then subjected for cytomorphometric and statistical analysis. Results: Assessment of quality using quality index revealed that PAP has better diagnostic efficiency as compared to MGG because the quality index of PAP was 0.87 and that of MGG was 0.74, the quality index of AO was 0.9, and that of DAPI was found to be 0.82, hence proving AO to be better in comparison with DAPI. Conclusion: We found that among all the four stains used AO is reliable, has greater efficacy, and can be easily performed with less cost. Despite the difficulties mentioned in this study, the progress in the field of fluorescence microscopy is expected to be rapid in the following years.