Spectral properties of fluorescence induced by glutaraldehyde fixation.
- 1 March 1981
- journal article
- research article
- Published by SAGE Publications in Journal of Histochemistry & Cytochemistry
- Vol. 29 (3), 411-414
- https://doi.org/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.This publication has 3 references indexed in Scilit:
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