The Histochemical Journal

, Volume 11, Issue 4, pp 473–483 | Cite as

Differentiation of the formaldehyde-induced fluorescent products of noradrenaline and dopamine by microspectrofluorometry

  • N. G. M. Wreford
  • G. C. Smith


The influence of concentration of dopamine and noradrenaline on the spectral characteristics of their formaldehyde-induced fluorophore, together with the influence of duration of reaction with formaldehyde, has been investigated in a model system. No substantial differences between fluorescence spectra of the amine fluorophores were observed. Accordingly, the influence of hydrochloric acid and ammonia vapours on the fluorophores was investigated. A shift to shorter wavelengths in the excitation maximum of each fluorophore was observed after a brief exposure to hydrochloric acid vapour; more prolonged exposure resulted in a pronounced shift back to longer wavelengths in the case of noradrenaline but no substantial change was observed with dopamine. Brief exposure to hydrochloric acid vapour resulted in a substantial increase in the rate of fading of the noradrenaline fluorophore on exposure to exciting radiation. It is suggested that this offers a convenient way of differentiating the amines.


Ammonia Formaldehyde Dopamine Noradrenaline Hydrochloric Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Björklund, A., Cegrell, L., Falck, B., Ritzén, M. &Rosengren, E. (1970). Dopamine-containing cells in sympathetic ganglia.Acta. physiol. scand. 78, 334–8.Google Scholar
  2. Björklund, A., Ehinger, B. &Falck, B. (1968). A method for differentiating dopamine from noradrenaline in tissue sections by microspectrofluorometry.J. Histochem. Cytochem. 16, 263–70.Google Scholar
  3. Björklund, A., Ehinger, B. &Falck, B. (1972). Analysis of fluorescence excitation peak ratios for the cellular identification of noradrenaline, dopamine or their mixtures.J. Histochem. Cytochem. 20, 56–64.Google Scholar
  4. Björklund, A. &Nobin, A. (1973). Fluorescence histochemical and microspectrofluorometric mapping of dopamine and noradrenaline cell groups in the rat diencephalon.Brain Res. 51, 193–205.Google Scholar
  5. Calas, A., Hartwig, H. G. &Collin, J. P. (1974). Noradrenergic innervation of the median eminence. Microspectrofluorometric and pharmacological study in the duckAnas platyrhynochos.Z. Zellforsch. 147, 491–504.Google Scholar
  6. Caspersson, T., Hillard, N. Å. &Ritzén, M. (1966). Fluorescence microspectrophotometry of cellular catecholamines and 5-hydroxytryptamine.Expl. Cell Res. 42, 415–28.Google Scholar
  7. Corrodi, H. &Jonsson, J. (1965). Fluorescence methods for the histochemical demonstration of monoamine: 4. Histochemical differentiation between dopamine and noradrenaline in models.J. Histochem. Cytochem. 13, 484–7.Google Scholar
  8. Corrodi, H., Jonsson, G. &Malmfors, T. (1966). Factors affecting the quality and intensity of the fluorescence in the histochemical method of demonstration of catecholamines.Acta histochem. 25, 367–70.Google Scholar
  9. Eränkö, O. (1961). Personal communication cited in Falck, B., Hillarp, N. A., Thieme, G. & Torp, A. (1962). Fluorescence, of catechol amines and related compounds condensed with formaldehyde.J. Histochem. Cytochem. 10, 348–54.Google Scholar
  10. Eränkö, O. &Raisanen, L. (1966).In vitro release and uptake, of noradrenaline in the rat iris. In:Mechanisms of Release of Biogenic Amines (Eds. U.S. von Euler, S. Rosel & B. Uvnas), pp. 73–78. Oxford: Pergamon.Google Scholar
  11. Eränkö, O. &Eränkö, L. (1971). Small intensely fluorescent granule-containing cells in sympathetic ganglion of the rat. InHistochemistry of Nervous Transmission. (Ed. O. Eränkö) Amsterdam: Elsevier.Google Scholar
  12. Ewen, S. W. B. &Rost, F. W. D. (1972). The histochemical demonstration of catecholamines and tryptamines by acid and aldehyde-induced fluorescence: microspectrofluorometric characterization of the fluorophores in models.Histochem. J. 4, 59–69.Google Scholar
  13. Falck, B. &Torp, A. (1961). A fluorescence method for the histochemical demonstration of noradrenaline in the adrenal medulla.Med. Exp. (Basel) 5, 428–32.Google Scholar
  14. Hamberger, B., Malmfors, T. &Sachs, C. (1965). Standardization of paraformaldehyde and of certain procedures for the histochemical demonstration of catecholamines.J. Histochem. Cytochem. 13, 147.Google Scholar
  15. Parker, C. A. &Rees, W. T. (1960). Correction of fluorescence spectra and measurement of fluorescence quantum efficiency.Analyst (Lond.) 85, 587–600.Google Scholar
  16. Ritzén, M. (1966). Quantitative fluorescence. Microspectrophotometry of catecholamine-formaldehyde products.Expl. Cell Res. 44, 505–20.Google Scholar
  17. Rybarczyk, K. E., Barker, H. A., Burke, J. P., Hartman, B. K. &Van Orden III, L. S. (1976). Histochemical and immunocytochemical identification of catecholamines, dopamine-β-hydroxylase and phenylethanolamine-N-methyl-transferase in S.I.F. cells. In:S.I.F. Cells: Structure and Function of the Small Intensely Fluorescent Sympathetic Cells. (Ed. O. Eränkö), pp. 68–81. Washington, D.C.: Department of Health Education and Welfare.Google Scholar
  18. Schofield, G. C. & Wreford, N. G. M. (1979). Microspectrofluorometric studies on the formaldehyde-induced reaction produced by 5-hydroxytryptamine.J. Microsc. (in press).Google Scholar
  19. Smith, G. C. &Helme, R. D. (1974). Ultrastructural and fluorescence histochemical studies on the effects of 6-hydroxydopamine on the rat median eminence.Cell Tiss. Res. 152, 493–512.Google Scholar
  20. Wreford, N. G. M. &Schofield, G. C. (1975). A microspectrofluorometer with on line real time correction of spectra.J. Microsc. 103, 127–30.Google Scholar
  21. Wreford, N. G. M. (1979). Microspectrofluorometry of cells in the mouse stomach.J. Anat. (in press).Google Scholar
  22. Van Orden, III, L. S. (1970). Quantitative histochemistry of biogenic amines. A simple microspectrofluorometer.Biochem. Pharmac. 19, 1105–17.Google Scholar

Copyright information

© Chapman and Hall Ltd. 1979

Authors and Affiliations

  • N. G. M. Wreford
    • 1
  • G. C. Smith
    • 1
  1. 1.Department of AnatomyMonash UniversityClaytonAustralia

Personalised recommendations