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This study introduces a novel Raman-microfluidic system for label-free cell sorting. The integrated system enables continuous, automated sorting of cyanobacteria with high purity using resonance Raman spectroscopy.

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Area of Science:

  • Biophotonics
  • Analytical Chemistry
  • Microfluidics

Background:

  • Single cell Raman spectroscopy offers label-free biochemical analysis of cells.
  • Weak intrinsic Raman signals pose challenges for discriminating and isolating cells in flow.
  • Existing methods often require cell trapping or chemical labeling.

Purpose of the Study:

  • To develop an integrated Raman-microfluidic system for continuous, label-free cell sorting.
  • To demonstrate automated sorting of cyanobacteria based on their biochemical fingerprints.
  • To overcome limitations of weak Raman signals and flow instability in microfluidic devices.

Main Methods:

  • An integrated Raman-microfluidic system utilizing "pressure dividers" for flow stabilization.
  • Resonance Raman spectroscopy to differentiate cyanobacteria cultured with (12)C or (13)C.
  • Real-time classification and sorting synchronized with Raman acquisition.
  • Continuous flow sorting without the need for cell trapping.

Main Results:

  • Stable flow profiles achieved using pressure dividers, enabling automated operation.
  • Successful sorting of cyanobacteria (Synechocystis sp. PCC6803) at low flow rates (<100 μm s(-1)).
  • Demonstrated sorting accuracy of 96.3% purity at a speed of 0.5 Hz.
  • Distinguished between (12)C and (13)C cultured microorganisms via subtle spectral differences.

Conclusions:

  • The developed Raman-microfluidic system provides a robust platform for label-free, continuous cell sorting.
  • Pressure dividers are crucial for stabilizing flow and enabling automated, high-purity sorting.
  • This technology has broad potential for analyzing and sorting various cell types based on their biochemical composition.