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We developed a mid-infrared photothermal imaging (MIPI) - fluorescence in-situ hybridization (FISH) microscope that enables simultaneous identity probing and metabolic activity imaging for bacteria in a complex environment. We added the 13C-glucose in the bacteria culture medium and monitored the newly synthesized protein composition by MIPI at amide I band. Bacterial taxa were targeted with fluorescently-labelled FISH probes and imaged with the widefield epi-fluorescence imaging that built on to the MIP microscope. The sub-micrometer spatial resolution of MIPI enables the characterization at single bacterium level. The proposed platform provides the link between genotype and phenotype and would be a powerful tool to deepen our understanding of microbiome.
Yeran Bai,Zhongyue Guo,Fátima Pereira,Michael Wagner, andJi-Xin Cheng
"Microbial identification and metabolic analysis by mid-infrared photothermal imaging fluorescence in-situ hybridization", Proc. SPIE PC11973, Advanced Chemical Microscopy for Life Science and Translational Medicine 2022, PC119730E (2 March 2022); https://doi.org/10.1117/12.2611781
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Yeran Bai, Zhongyue Guo, Fátima Pereira, Michael Wagner, Ji-Xin Cheng, "Microbial identification and metabolic analysis by mid-infrared photothermal imaging fluorescence in-situ hybridization," Proc. SPIE PC11973, Advanced Chemical Microscopy for Life Science and Translational Medicine 2022, PC119730E (2 March 2022); https://doi.org/10.1117/12.2611781