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Single cell analysis using surface enhanced Raman scattering (SERS) tags.

John P Nolan1, Erika Duggan, Er Liu

  • 1La Jolla Bioengineering Institute, 3535 General Atomics Court, San Diego, CA 92121, USA. jnolan@ljbi.org

Methods (San Diego, Calif.)
|April 14, 2012
PubMed
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Surface-enhanced Raman scattering (SERS) offers a powerful alternative to fluorescence for multiplexed bioanalysis. SERS-based flow cytometry enables high-content, multiparameter single-cell analysis with enhanced multiplexing capabilities.

Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Spectroscopy

Background:

  • Fluorescence is a common bioanalytical technique but has limitations in multiplexing due to broad emission spectra.
  • Conventional flow cytometry can measure many fluorophores but is limited by spectral overlap.
  • Surface-enhanced Raman scattering (SERS) offers narrow spectral features for increased multiplexing potential.

Purpose of the Study:

  • To develop nanoparticle SERS tags and Raman flow cytometers for multiparameter single-cell analysis.
  • To overcome the multiplexing limitations of fluorescence-based assays.
  • To enable high-content analysis of suspension and adherent cells.

Main Methods:

  • Development of plasmonically active gold nanorod SERS tags with tunable resonance.

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  • Adsorption of Raman resonant compounds onto nanoparticles for unique spectral fingerprints.
  • Encapsulation of SERS tags in polymer coatings for antibody conjugation.
  • Utilizing a high-resolution spectral flow cytometer for SERS and conventional measurements.
  • Application of automated spectral unmixing algorithms for data analysis.

    • Main Results:

    • SERS tags provide intense signals with narrow spectral features, enabling greater multiplexing.
    • Raman flow cytometry successfully measured complete SERS spectra from individual cells.
    • Automated spectral unmixing allowed extraction of individual SERS tag contributions.
    • High-content, multiparameter single-cell data was generated.

    Conclusions:

    • SERS-based cytometry is a powerful complement to fluorescence-based methods.
    • The narrow spectral features of SERS allow for higher levels of multiplexing with a single laser and detector.
    • This technology advances multiparameter single-cell analysis capabilities.