Highly multiplexed analysis of biospecimens significantly advances the understanding of biological basics of diseases, but these techniques are limited by the number of multiplexity and the speed of processing. Here, we present a rapid multiplex method for quantitative detection of protein markers on brain sections with the cellular resolution. This spatial multiplex in situ tagging (MIST) technology is built upon a MIST microarray that contains millions of small microbeads carrying barcoded oligonucleotides. Using antibodies tagged with UV cleavable oligonucleotides, the distribution of protein markers on a tissue slice could be “printed” on the MIST microarray with high fidelity. The performance of this technology in detection sensitivity, resolution and signal-to-noise level has been fully characterized by detecting brain cell markers. We showcase the codetection of 31 proteins simultaneously within 2 h which is about 10 times faster than the other immunofluorescence-based approaches of similar multiplexity. A full set of computational toolkits was developed to segment the small regions and identify the regional differences across the entire mouse brain. This technique enables us to rapidly and conveniently detect dozens of biomarkers on a tissue specimen, and it can find broad applications in clinical pathology and disease mechanistic studies.

中文翻译：

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用于快速、高度多路检测脑组织蛋白质分布的空间 MIST 技术

生物样本的高度多重分析显着促进了对疾病生物学基础的理解，但这些技术受到多重分析数量和处理速度的限制。在这里，我们提出了一种快速多重方法，用于定量检测具有细胞分辨率的脑切片上的蛋白质标记物。这种就地空间复用标记 (MIST) 技术建立在 MIST 微阵列之上，该微阵列包含数百万个携带条形码寡核苷酸的小微珠。使用带有 UV 可切割寡核苷酸标记的抗体，可以将组织切片上蛋白质标记的分布以高保真度“打印”在 MIST 微阵列上。该技术在检测灵敏度、分辨率和信噪比方面的性能已通过检测脑细胞标志物得到充分表征。我们展示了在 2 小时内同时对 31 种蛋白质进行共检测，这比其他基于免疫荧光的类似多重性方法快约 10 倍。开发了一整套计算工具包来分割小区域并识别整个小鼠大脑的区域差异。