2022/09/06 by Yeran Bai, Zhongyue Guo, Bai, Yeran +7
Biochemistry, Genetics and Molecular Biology · Engineering · Environmental Science · #Biological Physics (physics.bio-ph) #Biosensors and Analytical Detection #FOS: Biological sciences #FOS: Physical sciences #Genomics and Phylogenetic Studies #Microbial Community Ecology and Physiology #Quantitative Methods (q-bio.QM)
paper · pdf · doi:10.48550/arxiv.2209.03741
openalex publication_date 2022/09/06 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/28
Simultaneous identification and metabolic analysis of microbes with single-cell resolution and high throughput is necessary to answer the question of "who eats what, when, and where" in complex microbial communities. Here, we present a mid-infrared photothermal-fluorescence in situ hybridization (MIP-FISH) platform that enables direct bridging of genotype and phenotype. Through multiple improvements of MIP imaging, the sensitive detection of isotopically-labelled compounds incorporated into proteins of individual bacterial cells became possible, while simultaneous detection of FISH labelling with rRNA-targeted probes enabled the identification of the analyzed cells. In proof-of-concept experiments, we showed that the clear spectral red shift in the protein amide I region due to incorporation of 13C atoms originating from 13C-labelled-glucose can be exploited by MIP-FISH to discriminate and identify 13C-labelled bacterial cells within a complex human gut microbiome sample. The presented methods open new opportunities for single-cell structure-function analyses for microbiology.