Development overview of Raman-activated cell sorting devoted to bacterial detection at single-cell level

Understanding the metabolic interactions between bacteria in natural habitat at the single-cell level and the contribution of individual cell to their functions is essential for exploring the dark matter of uncultured bacteria. The combination of Raman-activated cell sorting (RACS) and single-cell Raman spectra (SCRS) with unique fingerprint characteristics makes it possible for research in the field of microbiology to enter the single cell era. This review presents an overview of current knowledge about the research progress of recognition and assessment of single bacterium cell based on RACS and further research perspectives. We first systematically summarize the label-free and non-destructive RACS strategies based on microfluidics, microdroplets, optical tweezers, and specially made substrates. The importance of RACS platforms in linking target cell genotype and phenotype is highlighted and the approaches mentioned in this paper for distinguishing single-cell phenotype include surface-enhanced Raman scattering (SERS), biomarkers, stable isotope probing (SIP), and machine learning. Finally, the prospects and challenges of RACS in exploring the world of unknown microorganisms are discussed.

Automated summary

Understanding metabolic interactions between bacteria at the single-cell level and the contribution of individual cells is crucial for exploring uncultured bacteria. The combination of RACS and SCRS with unique fingerprint characteristics has allowed microbiology research to enter the single-cell era. This review summarizes current research progress on recognizing and assessing single bacterial cells based on RACS, highlighting the importance of RACS platforms in linking cell genotype and phenotype, as well as methods for distinguishing single-cell phenotype.