Transcriptome-scale super-resolved imaging in tissues by RNA seqFISH
Transcriptome-scale super-resolved imaging in tissues by RNA seqFISH
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DOI：
10.1038/s41586-019-1049-y
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发表时间：
2019-04-11
期刊：
影响因子：
64.8
通讯作者：
Cai, Long
中科院分区：
文献类型：
--
作者：
Eng, Chee-Huat Linus;Lawson, Michael;Cai, Long
以高精度对转录组进行原位成像一直是单细胞生物学中的主要挑战，这尤其受到光学分辨率和单细胞中转录物密度的限制的阻碍（1-5）。在这里，我们展示了顺序荧光原位杂交（seqFISH+）的演变。我们表明，seqFISH+可以成像mRNA的10，000个基因在单细胞-具有高精度和子衍射极限分辨率-在皮质，脑室下区和嗅球的小鼠大脑，使用标准的共聚焦显微镜。seqFISH+的转录组水平分析允许无偏地鉴定细胞类别及其在组织中的空间组织。此外，seqFISH+揭示了细胞中的亚细胞mRNA定位模式和相邻细胞中的配体-受体对。该技术展示了生成空间细胞图谱和进行原位生物过程发现驱动研究的能力。
Imaging the transcriptome in situ with high accuracy has been a major challenge in single-cell biology, which is particularly hindered by the limits of optical resolution and the density of transcripts in single cells(1-5). Here we demonstrate an evolution of sequential fluorescence in situ hybridization (seqFISH+). We show that seqFISH+ can image mRNAs for 10,000 genes in single cells-with high accuracy and sub-diffraction-limit resolution-in the cortex, subventricular zone and olfactory bulb of mouse brain, using a standard confocal microscope. The transcriptome-level profiling of seqFISH+ allows unbiased identification of cell classes and their spatial organization in tissues. In addition, seqFISH+ reveals subcellular mRNA localization patterns in cells and ligand-receptor pairs across neighbouring cells. This technology demonstrates the ability to generate spatial cell atlases and to perform discovery-driven studies of biological processes in situ.