CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Nat Commun. 2023 Mar 17;14(1):1477. doi: 10.1038/s41467-023-37188-3.
Spatial analysis of microbiomes at single cell resolution with high multiplexity and accuracy has remained challenging. Here we present spatial profiling of a microbiome using sequential error-robust fluorescence in situ hybridization (SEER-FISH), a highly multiplexed and accurate imaging method that allows mapping of microbial communities at micron-scale. We show that multiplexity of RNA profiling in microbiomes can be increased significantly by sequential rounds of probe hybridization and dissociation. Combined with error-correction strategies, we demonstrate that SEER-FISH enables accurate taxonomic identification in complex microbial communities. Using microbial communities composed of diverse bacterial taxa isolated from plant rhizospheres, we apply SEER-FISH to quantify the abundance of each taxon and map microbial biogeography on roots. At micron-scale, we identify clustering of microbial cells from multiple species on the rhizoplane. Under treatment of plant metabolites, we find spatial re-organization of microbial colonization along the root and alterations in spatial association among microbial taxa. Taken together, SEER-FISH provides a useful method for profiling the spatial ecology of complex microbial communities in situ.
单细胞分辨率下高多重性和准确性的微生物组空间分析一直具有挑战性。在这里,我们使用顺序抗误差荧光原位杂交(SEER-FISH)来展示微生物组的空间分析,这是一种高度多重化和准确的成像方法,可实现微生物群落的微米级图谱。我们表明,通过探针杂交和解离的连续循环,可以显著提高微生物组中 RNA 分析的多重性。结合纠错策略,我们证明 SEER-FISH 可实现复杂微生物群落中的准确分类鉴定。使用来自植物根际的多种细菌分类群组成的微生物群落,我们应用 SEER-FISH 来定量每个分类群的丰度,并在根上绘制微生物生物地理学图谱。在微米尺度上,我们从根际上鉴定出多种微生物细胞的聚类。在植物代谢物处理下,我们发现微生物定殖沿根的空间重新组织以及微生物分类群之间空间关联的改变。总之,SEER-FISH 为原位分析复杂微生物群落的空间生态提供了一种有用的方法。