Portal de Periódicos da CAPES

Transcriptome-scale super-resolved imaging in tissues by RNA seqFISH+

2019; Nature Portfolio; Volume: 568; Issue: 7751 Linguagem: Inglês

10.1038/s41586-019-1049-y

1476-4687

Chee-Huat Linus Eng, Michael J. Lawson, Qian Zhu, Ruben Dries, Noushin Koulena, Yodai Takei, Jina Yun, Christopher J. Cronin, Christoph Karp, Guo‐Cheng Yuan, Long Cai,

Molecular Biology Techniques and Applications

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 cells1–5. 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. seqFISH+, an evolution of sequential fluorescence in situ hybridization with super-resolution imaging capabilities, is used to image mRNAs of 10,000 genes in cultured cells and mouse brain slices, demonstrating the ability to generate spatial atlases and to perform discovery-driven studies in situ.