空间转录组学能够实现组织中基因表达模式的单细胞分辨率视图,深入了解它们的生物学功能。然而,将这种方法应用于皮肤存在固有的挑战。这里,我们提出了一种用于制备包含毛囊的哺乳动物皮肤样品以进行空间转录组学的方案。我们描述了样品制备的步骤,嵌入,获取冷冻切片,RNA质量控制,组织安装,固定,染色,和成像。然后我们详细说明透化的程序,逆转录,和cDNA收集。有关此协议的使用和执行的完整详细信息,请参考Chen等人1。
Spatial transcriptomics enables a single-cell resolution view of gene expression patterns in tissues, providing insight into their biological functions. However, applying this approach to the skin presents inherent challenges. Here, we present a protocol for preparing mammalian skin samples encompassing hair follicles for spatial transcriptomics. We describe steps for sample preparation, embedding, acquisition of frozen slices, RNA quality control, tissue mounting, fixation, staining, and imaging. We then detail procedures for permeabilization, reverse transcription, and cDNA collection. For complete details on the use and execution of this protocol, please refer to Chen et al.1.