Abstract
Raman-activated cell sorting (RACS), which identifies and isolates cells according to their Raman spectra, would provide a powerful tool to explore uncultured bacteria in environment without cultivation. Single-cell Raman spectra (SCRS) are label-free, chemical profile of single cells, reflecting cell physiological states and metabolic activities. SCRS can be used as phenotypic signatures for individual cells. A combination of RACS and stable isotope probing (SIP) enables clearly distinguishing and separating of cells that perform relevant metabolic activities in environment. A strategy using controlled trap and release of single cell overcomes the problem of naturally weak Raman signals. This protocol provides the detailed description of RACS to isolate CO2-fixing photosynthetic cells in a microfluidic device. This protocol uses carotenoid-containing bacteria as an example for RACS; it can also be applied to other bacteria sorting by simply adjusting Raman acquisition time.
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We thank NERC (NE/M002934/1) for the fund support.
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Zhang, P., Song, Y., Thompson, I.P., Ma, B., Xu, J., Huang, W.E. (2015). Single-Cell Biotechnology for Uncultured Microorganisms. In: McGenity, T.J., Timmis, K.N., Nogales, B. (eds) Hydrocarbon and Lipid Microbiology Protocols. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8623_2015_116
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DOI: https://doi.org/10.1007/8623_2015_116
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