Nonlinear multicontrast microscopy of hematoxylin-and-eosin-stained histological sections

J Biomed Opt. 2010 Mar-Apr;15(2):026018. doi: 10.1117/1.3382908.

Abstract

Imaging hematoxylin-and-eosin-stained cancerous histological sections with multicontrast nonlinear excitation fluorescence, second- and third-harmonic generation (THG) microscopy reveals cellular structures with extremely high image contrast. Absorption and fluorescence spectroscopy together with second hyperpolarizability measurements of the dyes shows that strong THG appears due to neutral hemalum aggregation and is subsequently enhanced by interaction with eosin. Additionally, fluorescence lifetime imaging microscopy reveals eosin fluorescence quenching by hemalums, showing better suitability of only eosin staining for fluorescence microscopy. Multicontrast nonlinear microscopy has the potential to differentiate between cancerous and healthy tissue at a single cell level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms*
  • Contrast Media
  • Eosine Yellowish-(YS)*
  • Hematoxylin*
  • Image Enhancement / methods*
  • Image Interpretation, Computer-Assisted / methods*
  • Microscopy / methods*
  • Nonlinear Dynamics
  • Pattern Recognition, Automated / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Contrast Media
  • Eosine Yellowish-(YS)
  • Hematoxylin