Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-0054, Japan.
Department of Advanced Information Technology, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan.
Nat Commun. 2024 May 8;15(1):3657. doi: 10.1038/s41467-024-47989-9.
Cell states are regulated by the response of signaling pathways to receptor ligand-binding and intercellular interactions. High-resolution imaging has been attempted to explore the dynamics of these processes and, recently, multiplexed imaging has profiled cell states by achieving a comprehensive acquisition of spatial protein information from cells. However, the specificity of antibodies is still compromised when visualizing activated signals. Here, we develop Precise Emission Canceling Antibodies (PECAbs) that have cleavable fluorescent labeling. PECAbs enable high-specificity sequential imaging using hundreds of antibodies, allowing for reconstruction of the spatiotemporal dynamics of signaling pathways. Additionally, combining this approach with seq-smFISH can effectively classify cells and identify their signal activation states in human tissue. Overall, the PECAb system can serve as a comprehensive platform for analyzing complex cell processes.
细胞状态受信号通路对受体配体结合和细胞间相互作用的反应调节。高分辨率成像已被尝试用于探索这些过程的动态，最近，多重成像通过从细胞中全面获取空间蛋白质信息来描绘细胞状态。然而，在可视化激活信号时，抗体的特异性仍然受到影响。在这里，我们开发了具有可切割荧光标记的精确发射消除抗体（PECAbs）。PECAbs 可以使用数百种抗体进行高特异性的顺序成像，从而能够重建信号通路的时空动力学。此外，将这种方法与 seq-smFISH 相结合，可以有效地对人类组织中的细胞进行分类，并识别其信号激活状态。总的来说，PECAb 系统可以作为分析复杂细胞过程的综合平台。
