Translational Potential of Fluorescence Polarization for Breast Cancer Cytopathology

Simple Summary The evaluation of breast fine needle aspiration cytology specimens requires subjective, visual assessments of cytomorphology, resulting in suboptimal diagnostic accuracy. The fluorescence polarization of methylene blue demonstrated significant potential as a quantitative marker for cellular level breast cancer diagnosis in clinical aspirates. Results indicate the technology could be implemented as a standalone approach for breast cancer detection in singe cells or augment conventional approaches to reduce the incidence of indeterminate cytopathology. Breast cancer is the most common malignancy in women. The standard of care for diagnosis involves invasive core needle biopsy followed by time-consuming histopathological evaluation. A rapid, accurate, and minimally invasive method to diagnose breast cancer would be invaluable. Therefore, this clinical study investigated the fluorescence polarization (Fpol) of the cytological stain methylene blue (MB) for the quantitative detection of breast cancer in fine needle aspiration (FNA) specimens. Cancerous, benign, and normal cells were aspirated from excess breast tissues immediately following surgery. The cells were stained in aqueous MB solution (0.05 mg/mL) and imaged using multimodal confocal microscopy. The system provided MB Fpol and fluorescence emission images of the cells. Results from optical imaging were compared to clinical histopathology. In total, we imaged and analyzed 3808 cells from 44 breast FNAs. Fpol images displayed quantitative contrast between cancerous and noncancerous cells, whereas fluorescence emission images showed the morphological features comparable to cytology. Statistical analysis demonstrated that MB Fpol is significantly higher (p < 0.0001) in malignant vs. benign/normal cells. It also revealed a correlation between MB Fpol values and tumor grade. The results indicate that MB Fpol could provide a reliable, quantitative diagnostic marker for breast cancer at the cellular level.