Hu Xinyu, van Sluijs Bob, García-Blay Óscar, Huck Wilhelm T S, Hansen Maike M K
Institute for Molecules and Materials, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, the Netherlands; Oncode Institute, Nijmegen, the Netherlands.
Institute for Molecules and Materials, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, the Netherlands.
STAR Protoc. 2024 Dec 20;5(4):103336. doi: 10.1016/j.xpro.2024.103336. Epub 2024 Oct 1.
Understanding the molecular signatures of individual cells within complex biological systems is crucial for deciphering cellular heterogeneity and uncovering regulatory mechanisms. Here, we present a protocol for simultaneous multiplexed detection of selected mRNAs and (phospho-)proteins in mouse embryonic stem cells using spatial single-cell profiling. We describe steps for employing single-stranded DNA (ssDNA)-labeled antibo'dies, padlock probes, and rolling circle amplification to achieve simultaneous visualization of mRNAs and (phospho-)proteins at subcellular resolution. This protocol has potential application in identifying cells in heterogeneous biological microenvironments. For complete details on the use and execution of this protocol, please refer to Hu et al..
了解复杂生物系统中单个细胞的分子特征对于解读细胞异质性和揭示调控机制至关重要。在此，我们展示了一种使用空间单细胞分析同时多重检测小鼠胚胎干细胞中选定mRNA和（磷酸化）蛋白质的方案。我们描述了使用单链DNA（ssDNA）标记抗体、锁式探针和滚环扩增在亚细胞分辨率下实现mRNA和（磷酸化）蛋白质同时可视化的步骤。该方案在识别异质生物微环境中的细胞方面具有潜在应用。有关此方案使用和执行的完整详细信息，请参考Hu等人的研究。
