Selective staining by the fluorochrome, 5,5'-diphenyl-9-ethyl- DiOC2 (3). I. Physicochemical studies of dye-dye and dye-tissue interactions

Horst Hermel, Wolfgang Schmahl, Helmuth Möhwald

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The 5,5'-diphenyl-9-ethyl-oxacarbocyanine (5,5'-diphenyl-9-ethyl- DiOC2(3); CD) has properties suitable for histological investigations including the spectral range for absorption and fluorescence emission, the values of the corresponding molar coefficients and fluorescence quantum yield. Furthermore, CD remains relatively unchanged over the entire pH range and interacts with protein β-sheets. The latter fact is detectable spectroscopically as a bathochromic shift. In water-containing media such as the histological stain and washes, CD exists as a monomer, a dimer and in two aggregated states. These differ in their binding affinity to tissue sections, in their solubility in water, alcohol and water/alcohol mixtures, and in their UV/VIS absorption and fluorescence emission. The ratio of the various CD states and the contrast of selectively stained tissue areas can be controlled via the staining conditions and the sequence of the washes. Furthermore, mounting in a xylene-based medium produces a solvatochromic spectral shift of the CD monomer, which leads to a marked elevation in phase contrast.

Original languageEnglish
Pages (from-to)221-228
Number of pages8
JournalBiotechnic and Histochemistry
Volume74
Issue number5
DOIs
StatePublished - Sep 1999
Externally publishedYes

Keywords

  • Absorption
  • Fluorescence-emission
  • Oxacarbocyanine-dye
  • Protein β- sheets
  • Solvatochromism
  • Tissue-staining

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