High-speed hyperspectral imaging for highly multiplexed immunofluorescence imaging

项目摘要 使用免疫组织化学（IHC）的病理学仍然是现代临床医学中的重要工具。然而，在这方面， 常规免疫组化方法的能力尚未跟上生物医学的进步，这需要 评估网络，例如，在组织环境中的数十个大分子（通常是蛋白质靶点）， 多路复用为了克服这一限制，基于质谱和质谱分析的高度多路复用的IHC成像系统， 已经开发了重复免疫荧光（IF）成像。这些系统声称理论上 多路复用多达100个目标。然而，它们对于常规使用来说太慢、昂贵和复杂， 目前只在少数研究实验室使用。 我们正在努力从根本上改变IF成像的能力，以允许对数十个 分子靶向快速和适度的成本。我们的方法建立在我们以前的工作高速 高光谱成像结合放大分子靶标记方法。 我们将推进我们的技术，并通过具体的研究应用展示其能力。 涉及对来自与阿尔茨海默病相关的神经病理学小鼠模型的脑切片进行成像， （AD）。我们将进行改进，以实现更简单，更快地操作我们的技术。然后我们将 成像多达12个目标，并展示了将我们的方法扩展到单个组织上数十个目标的能力 滑盖.我们希望我们的仪器和协议将足够简单，低成本，快速，允许 大多数病理学和研究实验室的常规使用。 !

PROJECT SUMMARY Pathology using immunohistochemistry (IHC) remains a critical tool in modern clinical medicine. However, the capabilities of routine IHC methods have not kept pace with advances in biomedicine, which require evaluating networks of, e.g., tens of large molecules (typically protein targets) in a tissue context by multiplexing. To overcome this limit, highly multiplexed IHC imaging systems based on mass spectrometry and repeated Immunofluorescence (IF) imaging have been developed. These systems claim theoretical multiplexing of up to 100 targets. However, they are too slow, expensive, and complex, for routine use, and are currently used in a small number of research laboratories. We are working to radically change the capabilities of IF imaging to allow routine detection of tens of molecular targets rapidly and at moderate cost. Our approach builds on our previous work on high-speed hyperspectral imaging coupled with amplified molecular target labeling methods. We will advance our technology and demonstrate its capabilities with a specific research application involving imaging of brain sections from a mouse model of a neuropathology relevant for Alzheimer’s disease (AD). We will make improvements to achieve simpler and faster operation of our technology. We will then image up to 12 targets and demonstrate the ability to extend our approach to tens of targets on a single tissue slide. We expect our instrumentation and protocols will be sufficiently simple, low cost, and rapid to allow routine use in most pathology and research laboratories. !