ARUK-UCL Drug Discovery Institute, University College London, London, United Kingdom.
Department of Pathology and Molecular Pathology, University and University Hospital Zurich, Zurich, Switzerland.
Sci Rep. 2019 Aug 26;9(1):12353. doi: 10.1038/s41598-019-48787-w.
RNA analysis at the cellular resolution in the human brain is challenging. Here, we describe an optimised approach for detecting single RNA transcripts in a cell-type specific manner in frozen human brain tissue using multiplexed fluorescent RNAscope probes. We developed a new robust analytical approach for RNAscope quantification. Our method shows that low RNA integrity does not significantly affect RNAscope signal, recapitulates bulk RNA analysis and provides spatial context to transcriptomic analysis of human post-mortem brain at single-cell resolution. In summary, our optimised method allows the usage of frozen human samples from brain banks to perform quantitative RNAscope analysis.
在人类大脑中进行细胞分辨率的 RNA 分析具有挑战性。在这里，我们描述了一种优化的方法，用于使用多重荧光 RNAscope 探针以细胞类型特异性的方式检测冷冻人脑组织中的单个 RNA 转录本。我们开发了一种新的稳健的 RNAscope 定量分析方法。我们的方法表明，低 RNA 完整性不会显著影响 RNAscope 信号，可重现批量 RNA 分析，并为单细胞分辨率的人类死后大脑转录组分析提供空间背景。总之，我们的优化方法允许使用来自脑库的冷冻人类样本进行定量 RNAscope 分析。
