Spectral histopathology of colon cancer tissue sections by Raman imaging with 532 nm excitation provides label free annotation of lymphocytes, erythrocytes and proliferating nuclei of cancer cells

Analyst. 2013 Jul 21;138(14):4035-9. doi: 10.1039/c3an00370a. Epub 2013 Jun 4.

Abstract

Spectral histopathology (SHP) is an emerging tool for label free annotation of tissue. While FTIR based SHP provides fast annotation of larger tissue sections, Raman based SHP is slower but achieves a 10 times higher spatial resolution as compared to FTIR. Usually NIR excitation is used for Raman measurements on biological samples. Here, for the first time 532 nm excitation is used to annotate colon tissue by Raman SHP. Excellent data quality is obtained, which resolves for example erythrocytes and lymphocytes. In addition to Raman scattering auto-fluorescence is observed. We found that this auto-fluorescence overlaps spatially with the fluorescence of antibodies against p53 used in routine immunohistochemistry in surgical pathology. This fluorescence indicates nuclei of cancer cells with mutated p53 and allows new label free assignment of cancer cells. These results open new avenues for optical diagnosis by Raman spectroscopy and autofluorescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / genetics
  • Cell Nucleus / pathology*
  • Cell Proliferation*
  • Colonic Neoplasms / diagnosis*
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / surgery
  • Diagnostic Imaging*
  • Erythrocytes / pathology*
  • Fluorescence
  • Humans
  • Immunoenzyme Techniques
  • Lymphocytes / pathology*
  • Mutation / genetics
  • Spectroscopy, Fourier Transform Infrared
  • Spectrum Analysis, Raman / methods*
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • TP53 protein, human
  • Tumor Suppressor Protein p53