Chakraborty Sandeep, Chen Sheng-Tse, Hsiao Yang-Ting, Chiu Ming-Jang, Sun Chi-Kuang
Department of Electrical Engineering and Graduate Institute of Photonics and Optoelectronics, National Taiwan University, Taipei 10617, Taiwan.
Department of Neurology, National Taiwan University Hospital, Taipei 10051, Taiwan.
Biomed Opt Express. 2020 Jan 2;11(2):571-585. doi: 10.1364/BOE.378447. eCollection 2020 Feb 1.
Multicolor fluorescence imaging has been widely used by neuroscientists to simultaneously observe different neuropathological features of the brain. However, these optical modalities rely on exogenous labeling. Here, we demonstrate, for the first time, a label-free additive-color multi-harmonic generation microscopy to elucidate, concurrently with different hues, Alzheimer's disease (AD) neuropathological hallmarks: amyloid (A) plaques and neurofibrillary tangles (NFT). By treating third harmonic generation (THG) and second harmonic generation (SHG) as two primary colors, our study can simultaneously label-free differentiate AD hallmarks by providing different additive colors between A plaques, NFT, and neuronal axons, with weaker THG presentation from NFT in most places of the brain. Interestingly our pixel-based quantification and Pearson's correlation results further corroborated these findings. Our proposed label-free technique fulfills the unmet challenge in the clinical histopathology for stain-free slide-free differential visualization of neurodegenerative disease pathologies, with a sub-femtoliter resolution in a single image field-of-view.
多色荧光成像已被神经科学家广泛用于同时观察大脑的不同神经病理学特征。然而，这些光学模态依赖于外源性标记。在此，我们首次展示了一种无标记的加色多谐波产生显微镜，能够与不同色调同时阐明阿尔茨海默病（AD）的神经病理学特征：淀粉样蛋白（A）斑块和神经原纤维缠结（NFT）。通过将三次谐波产生（THG）和二次谐波产生（SHG）视为两种原色，我们的研究能够通过在A斑块、NFT和神经元轴突之间提供不同的加色，以无标记方式同时区分AD特征，在大脑的大多数部位，NFT的THG呈现较弱。有趣的是，我们基于像素的量化和皮尔逊相关性结果进一步证实了这些发现。我们提出的无标记技术满足了临床组织病理学中对神经退行性疾病病理进行无染色、无切片差异可视化的未满足挑战，在单个图像视野中具有亚飞升分辨率。
