University of Massachusetts Lowell, Advanced Biophotonics Laboratory, 175 Cabot Street, Suite 110-111, Lowell, Massachusetts 01854, USA.
J Biomed Opt. 2012 Jun;17(6):066008. doi: 10.1117/1.JBO.17.6.066008.
The goal of the study was to evaluate wide-field and high-resolution multimodal optical imaging, including polarization, reflectance, and fluorescence for the intraoperative detection of breast cancer. Lumpectomy specimens were stained with 0.05 mg/ml aqueous solution of methylene blue (MB) and imaged. Wide-field reflectance images were acquired between 390 and 750 nm. Wide-field fluorescence images were excited at 640 nm and registered between 660 and 750 nm. High resolution confocal reflectance and fluorescence images were excited at 642 nm. Confocal fluorescence images were acquired between 670 nm and 710 nm. After imaging, the specimens were processed for hematoxylin and eosin (H&E) histopathology. Histological slides were compared with wide-field and high-resolution optical images to evaluate correlation of tumor boundaries and cellular morphology, respectively. Fluorescence polarization imaging identified the location, size, and shape of the tumor in all the cases investigated. Averaged fluorescence polarization values of tumor were higher as compared to normal tissue. Statistical analysis confirmed the significance of these differences. Fluorescence confocal imaging enabled cellular-level resolution. Evaluation and statistical analysis of MB fluorescence polarization values registered from single tumor and normal cells demonstrated higher fluorescence polarization from cancer. Wide-field high-resolution fluorescence and fluorescence polarization imaging shows promise for intraoperative delineation of breast cancers.
本研究旨在评估宽场和高分辩率多模态光学成像技术,包括偏振、反射和荧光,以实现乳腺癌的术中检测。切除的标本用 0.05mg/ml 的亚甲蓝(MB)水溶液染色,并进行成像。采集 390nm 至 750nm 之间的宽场反射图像。用 640nm 激发宽场荧光图像,并在 660nm 至 750nm 之间进行登记。用 642nm 激发高分辨率共聚焦反射和荧光图像。共聚焦荧光图像在 670nm 至 710nm 之间采集。成像后,标本进行苏木精和伊红(H&E)组织病理学处理。将组织学幻灯片与宽场和高分辨率光学图像进行比较,分别评估肿瘤边界和细胞形态的相关性。荧光偏振成像在所有研究的病例中均能识别肿瘤的位置、大小和形状。与正常组织相比,肿瘤的平均荧光偏振值更高。统计分析证实了这些差异的显著性。荧光共聚焦成像实现了细胞级分辨率。对来自单个肿瘤和正常细胞的 MB 荧光偏振值的评估和统计分析表明,来自癌症的荧光偏振值更高。宽场高分辨率荧光和荧光偏振成像有望用于乳腺癌的术中描绘。