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An automated Raman-based platform for the sorting of live cells by functional properties

Research output: Contribution to journalArticle

  • Kang Soo Lee
  • Márton Palatinszky
  • Fatima C. Pereira
  • Jen Nguyen
  • Vicente Fernandez
  • Anna Mueller
  • Filippo Menolascina
  • Holger Daims
  • David Berry
  • Michaela Wagner
  • Roman Stocker

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https://www.nature.com/articles/s41564-019-0394-9
Original languageEnglish
JournalNature Microbiology
Early online date18 Mar 2019
DOIs
Publication statusE-pub ahead of print - 18 Mar 2019

Abstract

Stable-isotope probing is widely used to study the function of microbial taxa in their natural environment, but sorting of isotopically labelled microbial cells from complex samples for subsequent genomic analysis or cultivation is still in its early infancy. Here, we introduce an optofluidic platform for automated sorting of stable-isotope-probing-labelled microbial cells, combining microfluidics, optical tweezing and Raman microspectroscopy, which yields live cells suitable for subsequent single-cell genomics, mini-metagenomics or cultivation. We describe the design and optimization of this Raman-activated cell-sorting approach, illustrate its operation with four model bacteria (two intestinal, one soil and one marine) and demonstrate its high sorting accuracy (98.3 ± 1.7%), throughput (200–500 cells h−1; 3.3–8.3 cells min−1) and compatibility with cultivation. Application of this sorting approach for the metagenomic characterization of bacteria involved in mucin degradation in the mouse colon revealed a diverse consortium of bacteria, including several members of the underexplored family Muribaculaceae, highlighting both the complexity of this niche and the potential of Raman-activated cell sorting for identifying key players in targeted processes.

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