Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Gubra ApS, Hørsholm, Denmark.
Science. 2024 Nov 22;386(6724):907-915. doi: 10.1126/science.adn9947. Epub 2024 Nov 21.
Recent advances in RNA analysis have deepened our understanding of cellular states in biological tissues. However, a substantial gap remains in integrating RNA expression data with spatial context across organs, primarily owing to the challenges associated with RNA detection within intact tissue volumes. Here, we developed Tris buffer-mediated retention of in situ hybridization chain reaction signal in cleared organs (TRISCO), an effective tissue-clearing method designed for whole-brain spatial three-dimensional (3D) RNA imaging. TRISCO resolved several crucial issues, including the preservation of RNA integrity, achieving uniform RNA labeling, and enhancing tissue transparency. We tested TRISCO using a broad range of cell-identity markers, noncoding and activity-dependent RNAs, within diverse organs of varying sizes and species. TRISCO thus emerges as a powerful tool for single-cell, whole-brain, 3D imaging that enables comprehensive transcriptional spatial analysis across the entire brain.
RNA 分析的最新进展加深了我们对生物组织中细胞状态的理解。然而，由于在完整组织体积内检测 RNA 相关的挑战，将 RNA 表达数据与器官的空间背景进行整合仍然存在很大差距。在这里，我们开发了 Tris 缓冲液介导的原位杂交链反应信号在透明化组织中的保留（TRISCO），这是一种针对全脑空间三维（3D）RNA 成像设计的有效的组织透明化方法。TRISCO 解决了几个关键问题，包括 RNA 完整性的保留、实现均匀的 RNA 标记和增强组织透明度。我们使用广泛的细胞身份标记物、非编码 RNA 和活性依赖型 RNA 在不同大小和物种的各种器官中测试了 TRISCO。因此，TRISCO 成为一种用于单细胞、全脑、3D 成像的强大工具，可实现整个大脑的全面转录空间分析。
