Raman-Activated, Interactive Sorting of Isotope-Labeled Bacteria

Area of Science:

• Microbiology
• Spectroscopy
• Biotechnology

Background:

• Raman microspectroscopy offers high spatial resolution for analyzing single microbial cells and their metabolic states using stable isotope probing (SIPs).
• Static Raman measurements are limited as they do not allow for subsequent analysis of the analyzed cells.
• Combining Raman analysis with cell sorting is crucial for downstream research on selected microbial populations.

Purpose of the Study:

• To develop a novel microfluidic approach for Raman-activated continuous-flow sorting of bacteria.
• To integrate image-based particle sorting with synchronous Raman spectral acquisition and analysis for real-time sorting decisions.
• To enable the isolation of metabolically active microbial cells for further investigation.

Main Methods:

• Development of a microfluidic chip for continuous-flow cell sorting.
• Implementation of an optical setup for image-based particle sorting.
• Synchronous acquisition and analysis of Raman spectra to inform sorting decisions.
• Demonstration of Raman-activated sorting of microbial cells.

Main Results:

• Successful demonstration of a microfluidic approach for Raman-activated cell sorting.
• The system enables sorting based on Raman spectral information, reflecting cellular content and metabolic state.
• Active microbial cells were successfully sorted using the developed microfluidic chip and optical setup.

Conclusions:

• The developed microfluidic chip provides a powerful tool for isolating specific microbial cells based on their phenotypic and metabolic characteristics.
• This technology facilitates downstream analysis of sorted cells, opening new avenues for microbial research.
• Raman-activated sorting represents a significant advancement in microbial cell analysis and isolation techniques.