University of Toronto, Department of Physics, Institute for Optical Sciences, Department of Chemical and Physical Sciences, 3359 Mississauga Road North, Mississauga, Ontario, L5L 1C6 Canada.
J Biomed Opt. 2010 Mar-Apr;15(2):026018. doi: 10.1117/1.3382908.
Imaging hematoxylin-and-eosin-stained cancerous histological sections with multicontrast nonlinear excitation fluorescence, second- and third-harmonic generation (THG) microscopy reveals cellular structures with extremely high image contrast. Absorption and fluorescence spectroscopy together with second hyperpolarizability measurements of the dyes shows that strong THG appears due to neutral hemalum aggregation and is subsequently enhanced by interaction with eosin. Additionally, fluorescence lifetime imaging microscopy reveals eosin fluorescence quenching by hemalums, showing better suitability of only eosin staining for fluorescence microscopy. Multicontrast nonlinear microscopy has the potential to differentiate between cancerous and healthy tissue at a single cell level.
用多对比度非线性激发荧光对苏木精和曙红染色的癌症组织切片进行成像，二次和三次谐波产生（THG）显微镜可以揭示具有极高图像对比度的细胞结构。染料的吸收和荧光光谱以及二阶超极化率测量表明，强烈的 THG 是由于中性血晶聚集引起的，随后与曙红相互作用而增强。此外，荧光寿命成像显微镜揭示了血晶对曙红荧光的猝灭，表明只有曙红染色更适合荧光显微镜。多对比度非线性显微镜有可能在单细胞水平上区分癌症组织和健康组织。
