SRS-FISH: high-throughput bridging of phenotype and genotype at single cell level

Xiaowei Ge; Fatima C. Pereira; Meng Zhang; Peng Lin; Michael Wagner; Ji-Xin Cheng

doi:10.1117/12.2577969

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5 March 2021 SRS-FISH: high-throughput bridging of phenotype and genotype at single cell level

Xiaowei Ge, Fatima C. Pereira, Meng Zhang, Peng Lin, Michael Wagner, Ji-Xin Cheng

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Proceedings Volume 11656, Advanced Chemical Microscopy for Life Science and Translational Medicine 2021; 116560W (2021) https://doi.org/10.1117/12.2577969
Event: SPIE BiOS, 2021, Online Only

Abstract

For high-throughput link of microbiome function and taxonomic identity at the single cell level, we established a stimulated Raman scattering (SRS)-fluorescence in situ hybridization (FISH) platform. SRS combined with the deuterium-based isotope probing enables chemical mapping and reveals metabolic activity of bacteria. Fluorescently tagged oligonucleotide probes identify different bacteria and are detected through two photon fluorescence (TPF) microscopy. As a proof-of-principle demonstration, we tested the platform in a mixture of two distinct gut microbiota taxa with different deuterium labeling levels. This established platform not only provides enormous potential to study microbiota in the complex environment, but also the simultaneous observation of phenotype and genotype in the general biological systems.

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Xiaowei Ge, Fatima C. Pereira, Meng Zhang, Peng Lin, Michael Wagner, and Ji-Xin Cheng "SRS-FISH: high-throughput bridging of phenotype and genotype at single cell level", Proc. SPIE 11656, Advanced Chemical Microscopy for Life Science and Translational Medicine 2021, 116560W (5 March 2021); https://doi.org/10.1117/12.2577969

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