Spectral properties of fluorescence induced by glutaraldehyde fixation

J Histochem Cytochem. 1981 Mar;29(3):411-4. doi: 10.1177/29.3.6787116.

Abstract

Following fixation with glutaraldehyde, tissues or gelatin films fluoresce. This fluorescence can be enhanced more than thirtyfold by several minutes exposure to near ultraviolet light. Longer wavelengths produce a smaller effect. The enhanced fluorescence is maximally excited at 540 nm (half-band width about 45 nm), and fluorescence emission peaks at 560 nm. The rate of photoenhancement is independent of temperature in the 12--30 degrees C range. Photoenhancement is greater at alkaline than acid pH; the pH dependence involves a single acid binding group with pK = 7.3. These effects are not observed following treatment with paraformaldehyde or prior reduction with boranedimethylamine. Schiff base linkages between the bifunctional cross-linking reagent and free amino groups therefore seem to be involved. The effects of pH and wavelength on the photoenhancement of fluorescence are best accounted for by a kinetic scheme that includes both photogeneration of fluorophore from an ultraviolet-absorbing precursor, and its subsequent photodestruction by visible light.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aldehydes*
  • Animals
  • Astacoidea
  • Cell Membrane / ultrastructure*
  • Glutaral*
  • Indicators and Reagents*
  • Microscopy, Fluorescence / methods*
  • Microvilli / ultrastructure*
  • Ultraviolet Rays

Substances

  • Aldehydes
  • Indicators and Reagents
  • Glutaral