Comparative Analysis of Eosin-based Fluorescence Microscopy of Non-neoplastic Breast Tissue and Fibroadenoma

Abstract

Background: Fibroadenoma (FA) is one of the most frequently diagnosed benign neoplasm in women. Various researches have reported increased risk of breast cancer in females with FA. It stems from the proliferation of epithelial and stromal contents of the terminal duct lobular units (TDLU`S) of breast tissue, that are the primary sites for the histopathologic assessment which is the gold standard for the diagnosis of disease. However, this method is subjective and possess interobserver variability. Therefore, new quantitative methods are required to aid in diagnosis. Hence we evaluated fluorescence light intensity and its use in histopatholgic evaluation. Aim: The goal of this research was to compare and quantify red and green fluorescence light intensities of ductal cells and stroma of non-neoplastic breast tissue with fibroadenomatous tissue. Method: A cross-sectional study was done in the Cell biology and histology lab of Ziauddin University. 44 slides of normal breast tissue and 44 slides of diagnosed fibroadenomatous tissue were taken from Dr Ziauddin Hospital, North Campus. Hematoxylin and eosin (H&E) staining of the slides were done following standard protocols. On microscopic examination, the changes in light intensities of ductal cells and stroma of normal breast tissue and fibroadenoma were quantified using dual channel fluorescence microscopy using Nikon NIS imaging software. Results: The results demonstrated statistically significant increase (p-value <0.05) in mean red (37.22±5.9) and green (22.47±6.6)  light intensity of stroma in FA when compared with red (32.71± 6.7) and green (17.01±4.3) light intensity of normal breast tissue. Whereas, R/G ratio for normal tissue was higher (1.95±0.11) than R/G for FA (1.74±0.37) with a p value of <0.05. Similarly, for ductal cells; statistically significant (p value <0.05) increase in mean red (38.86±5.4) and green (15.54±2.51) light intensity for FA was found when compared with red (29.62±1.89) and green (12.60±1.67) intensity of normal tissue. R/G ratio for FA (2.5±0.24) was compared to be higher than normal tissue (2.36±0.3) with a p value of <0.05. Conclusion: The study suggests that fluorescence microscopy combined with quantitative assessment fluorescence light intensities may may be a helpful tool for histomorphic evaluation of the breast tissue.