Immuno-surface-enhanced coherent anti-stokes Raman scattering microscopy: immunohistochemistry with target-specific metallic nanoprobes and nonlinear Raman microscopy

Anal Chem. 2011 Sep 15;83(18):7081-5. doi: 10.1021/ac201284d. Epub 2011 Aug 26.

Abstract

Immunohistochemistry (IHC) is one of the most widely used staining techniques for diagnostic purposes. The selective localization of target proteins in tissue specimens by conventional IHC is achieved with dye- or enzyme-labeled antibodies in combination with light microscopy. In this contribution, we demonstrate the proof-of-principle for IHC based on surface-enhanced coherent Raman scattering for contrast generation. Specifically, antibody-labeled metallic nanoshells in conjunction with surface-enhanced coherent anti-Stokes Raman scattering (SECARS) microscopy are employed for the selective, sensitive, and rapid localization of the basal cell protein p63 in normal prostate tissue. Negative control experiments were performed in order to confirm the selective binding of the target-specific metal nanoprobes and to disentangle the role of plasmonic (metal) and molecular (Raman reporter) resonances in this plasmon-assisted four-wave mixing technique.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Immobilized / chemistry
  • Antibodies, Immobilized / immunology
  • Gold / chemistry
  • Humans
  • Immunohistochemistry
  • Male
  • Metal Nanoparticles / chemistry*
  • Microscopy / methods*
  • Prostate / metabolism
  • Prostate / pathology
  • Silver / chemistry
  • Spectrum Analysis, Raman / methods*
  • Transcription Factors / analysis
  • Transcription Factors / immunology
  • Tumor Suppressor Proteins / analysis
  • Tumor Suppressor Proteins / immunology

Substances

  • Antibodies, Immobilized
  • TP63 protein, human
  • Transcription Factors
  • Tumor Suppressor Proteins
  • Silver
  • Gold