Translational Potential of Fluorescence Polarization for Breast Cancer Cytopathology.

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.

乳腺细针穿刺细胞学标本的评估需要对细胞形态进行主观的目视评估，这导致诊断准确性欠佳。亚甲蓝的荧光偏振在临床穿刺样本中作为细胞水平乳腺癌诊断的定量标志物显示出巨大潜力。结果表明，该技术可作为一种独立的单细胞乳腺癌检测方法，或者可增强常规方法以减少不确定细胞病理学的发生率。 乳腺癌是女性中最常见的恶性肿瘤。诊断的标准方法包括有创的粗针活检，随后是耗时的组织病理学评估。一种快速、准确且微创的乳腺癌诊断方法将具有极大的价值。因此，这项临床研究探究了细胞染色剂亚甲蓝（MB）的荧光偏振（Fpol）在细针穿刺（FNA）标本中对乳腺癌的定量检测。手术后立即从多余的乳腺组织中抽取癌性、良性和正常细胞。这些细胞在亚甲蓝水溶液（0.05毫克/毫升）中染色，并使用多模式共聚焦显微镜成像。该系统提供了细胞的亚甲蓝荧光偏振和荧光发射图像。光学成像的结果与临床组织病理学进行了比较。我们总共对来自44例乳腺细针穿刺的3808个细胞进行了成像和分析。荧光偏振图像显示了癌细胞与非癌细胞之间的定量对比，而荧光发射图像显示出与细胞学相当的形态特征。统计分析表明，恶性细胞与良性/正常细胞相比，亚甲蓝荧光偏振显著更高（p < 0.0001）。它还揭示了亚甲蓝荧光偏振值与肿瘤分级之间的相关性。结果表明，亚甲蓝荧光偏振可在细胞水平为乳腺癌提供一种可靠的定量诊断标志物。