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Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
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Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering.

Yuta Suzuki1, Koya Kobayashi1, Yoshifumi Wakisaka2

  • 1Department of Electrical Engineering and Information Systems, The University of Tokyo, 113-8656 Tokyo, Japan.

Proceedings of the National Academy of Sciences of the United States of America
|July 21, 2019
PubMed
Summary

This study introduces label-free chemical imaging flow cytometry using stimulated Raman scattering (SRS) microscopy. This breakthrough enables high-throughput, marker-free analysis of cellular metabolism and cancer detection without fluorescent labels.

Keywords:
cancer cellsimaging flow cytometrymetabolite imagingmicroalgaestimulated Raman scattering

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Area of Science:

  • Biotechnology and Biomedical Engineering
  • Chemical Imaging and Spectroscopy
  • Cellular Biology and Metabolism

Background:

  • Imaging flow cytometry is a powerful tool for analyzing cellular phenotypes but relies on fluorescent labeling.
  • The requirement for fluorescent labels limits its application in certain biological and clinical studies.
  • Developing label-free methods is crucial for expanding the utility of high-throughput cell analysis.

Purpose of the Study:

  • To develop and demonstrate a label-free chemical imaging flow cytometry technique.
  • To achieve high-throughput, multicolor stimulated Raman scattering (SRS) microscopy of flowing cells.
  • To showcase the application of this technology in studying cellular metabolism and cancer detection.

Main Methods:

  • Integration of a pulse pair-resolved wavelength-switchable Stokes laser with SRS microscopy.
  • Utilized a 3D acoustic focusing microfluidic chip for precise cell manipulation.
  • Employed deep learning algorithms for data analysis and interpretation.

Main Results:

  • Achieved unprecedented throughput of up to ~140 cells/s for multicolor SRS imaging of flowing cells.
  • Successfully studied metabolic heterogeneity in microalgal cells without fluorescent markers.
  • Demonstrated marker-free cancer detection in blood samples using label-free chemical imaging.

Conclusions:

  • Label-free chemical imaging flow cytometry overcomes the limitations of fluorescent labeling.
  • The developed SRS imaging flow cytometry offers high throughput and broad applicability.
  • This technology holds significant potential for advancing cellular analysis in research and diagnostics.