Simple, Scalable Proteomic Imaging for High-Dimensional Profiling of Intact Systems.

Combined measurement of diverse molecular and anatomical traits that span multiple levels remains a major challenge in biology. Here, we introduce a simple method that enables proteomic imaging for scalable, integrated, high-dimensional phenotyping of both animal tissues and human clinical samples. This method, termed SWITCH, uniformly secures tissue architecture, native biomolecules, and antigenicity across an entire system by synchronizing the tissue preservation reaction. The heat- and chemical-resistant nature of the resulting framework permits multiple rounds (>20) of relabeling. We have performed 22 rounds of labeling of a single tissue with precise co-registration of multiple datasets. Furthermore, SWITCH synchronizes labeling reactions to improve probe penetration depth and uniformity of staining. With SWITCH, we performed combinatorial protein expression profiling of the human cortex and also interrogated the geometric structure of the fiber pathways in mouse brains. Such integrated high-dimensional information may accelerate our understanding of biological systems at multiple levels.

对跨越多个层面的多种分子和解剖特征进行联合测量仍然是生物学中的一项重大挑战。在此，我们介绍一种简单的方法，该方法能够实现蛋白质组学成像，用于对动物组织和人类临床样本进行可扩展的、综合的高维表型分析。这种被称为SWITCH的方法通过同步组织保存反应，在整个系统中均匀地固定组织结构、天然生物分子和抗原性。所形成的框架具有耐热和耐化学性，允许进行多轮（>20轮）重新标记。我们对单个组织进行了22轮标记，并对多个数据集进行了精确的配准。此外，SWITCH使标记反应同步，以提高探针穿透深度和染色的均匀性。利用SWITCH，我们对人类大脑皮层进行了组合蛋白质表达谱分析，还探究了小鼠大脑中纤维通路的几何结构。这种综合的高维信息可能会加速我们对多个层面生物系统的理解。