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High-throughput label-free molecular fingerprinting flow cytometry.

Kotaro Hiramatsu1,2,3, Takuro Ideguchi2,3,4, Yusuke Yonamine5

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We developed a label-free Raman flow cytometer for high-throughput single-cell analysis. This technology chemically probes cells, overcoming limitations of fluorescent labeling and enabling new biological insights.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Cell Biology

Background:

  • Flow cytometry is crucial for analyzing cell populations but relies on fluorescent labels.
  • Fluorescent labeling has inherent limitations and can affect cell viability.
  • Label-free methods offer an alternative for cell analysis without exogenous markers.

Purpose of the Study:

  • To introduce a novel high-throughput Raman flow cytometer for label-free single-cell analysis.
  • To demonstrate the capability of chemically probing single live cells using molecular vibrational spectroscopy.
  • To showcase a practical application in analyzing astaxanthin productivity and photosynthetic dynamics.

Main Methods:

  • Development of a microfluidic chip-based Raman flow cytometer.
  • Utilizing a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering (FT-CARS) spectrometer.
  • Acquisition of broadband molecular vibrational spectra from single cells at ~2000 events/s.

Main Results:

  • Successful implementation of label-free chemical probing of single live cells.
  • Achieved high-throughput spectral acquisition in the fingerprint region (400–1600 cm⁻¹).
  • Demonstrated label-free analysis of astaxanthin production and photosynthesis in *Haematococcus lacustris*.

Conclusions:

  • The developed Raman flow cytometer offers a powerful label-free alternative to conventional methods.
  • This technology enables high-throughput analysis of cellular chemistry and dynamics.
  • Opens new avenues for studying biological processes without fluorescent interference.