High-speed hyperspectral imaging for highly multiplexed immunofluorescence imaging

项目总结 免疫组织化学病理学(IHC)在现代临床医学中仍然是一个重要的工具。然而， 常规IHC方法的能力没有跟上生物医学的进步，这需要 在组织环境中评估例如数十个大分子(通常是蛋白质靶标)的网络 多路传输。为了克服这一限制，基于质谱学和 重复免疫荧光(IF)成像已经发展起来。这些系统声称理论上 多路传输多达100个目标。然而，对于常规使用来说，它们太慢、太贵、太复杂，而且 目前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 分子靶标迅速，成本适中。我们的方法建立在我们以前在高速 高光谱成像与放大分子靶标记法相结合。 我们将推进我们的技术，并通过具体的研究应用来展示其能力 涉及与阿尔茨海默病相关的神经病理的小鼠模型的脑切片成像 (Ad)。我们将进行改进，以实现我们的技术更简单、更快的操作。到时候我们会的 成像多达12个靶点，并展示将我们的方法扩展到单个组织上数十个靶点的能力 滑行。我们希望我们的仪器和协议足够简单、低成本和快速，以允许 大多数病理学和研究实验室的常规使用。 好了！

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. !