Summary
This paper describes new procedures for highly sensitive visualization of monoamine stores in peripheral tissues, taking advantage of the recently introduced aluminum-catalysed formaldehyde (ALFA) reaction. The tissues are exposed to an aluminum sulphate solution (with or without formaldehyde fixation) in a perfusion and/or immersion step, followed by formaldehyde vapour treatment. Procedures are described for freeze-dried, paraffin embedded tissue, cryostat sections and whole mount preparations. For all these tissue preparations the ALFA method gives a highly sensitive and precise demonstration of catecholamine-containing neurons and 5-HT-containing cells in a variety of peripheral tissues. For freeze-dried tissue and cryostat sections the ALFA method represents an improvement in comparison with other available methods. This is particularly noticeable for the very delicate adrenergic nerves in such organs as the thyroid, ovary, pancreas and the gastrointestinal tract.
Similar content being viewed by others
References
Alm, P., Cegrell, L., Ehinger, B., Falck, B.: Remarkable adrenergic nerves in the exocrine pancreas. Z. Zellforsch. 83, 178–186 (1967)
Alm, P., Liedberg, G., Owman, Ch.: Gastric and pancreatic sympathetic denervation in the rat. Scand. J. Gastroenterol. 6, 307–312 (1971)
Battenberg, E.L.F., Bllom, F.E.: A rapid, simple and more sensitive method for the demonstration of central catecholamine-containing neurons and axons by glyoxylic acid induced fluorescence. I. Specificity. Psychopharmacol. Commun. I, 3–13 (1975)
Björklund, A., Falck, B., Owman, Ch.: Fluorescence microscopic and microspectrofluoremetric techniques for the cellular localization and characterization of biogenic amines. In: Methods of investigative and diagnostic endocrinology. S.A. Berson (ed). Vol. I: The theyroid and biogenic amines J.E. Rall and I.J. Kopin (eds.), pp. 318–368. Amsterdam: North-Holland 1972
Björklund, A., Falck, B., Lindvall, O., Lorén, I.: The aluminum-formaldehyde (ALFA) histofluorescence method for improved visualization of catecholamines and indolamines. II. Model experiments. J. Neurosci. Methods, in press (1980)
Bloom, F.E., Battenberg, E.L.F.: A rapid, simple and more sensitive method for the demonstration of central catecholamine-containing neurons and axons by glyoxylic acid induced fluorescence: II: A detailed description of methodology. J. Histochem. Cytochem. 24, 561–571 (1976)
Cegrell, L.: The occurrence of biogenic monoamines in the mammalian endocrine pancreas. Acta Physiol. Scand., Suppl. 314 (1968)
Corrodi, H., Hillarp, N.-Å.: Fluoreszenzmethoden zur histochemischen Sichtbarmachung von Monoaminen. I. Identifizierung der fluoreszierenden Produkte aus Modellversuchen mit 6,7-Dimethoxyisochinolinderivaten und Formaldehyd. Helv. Chim. Acta 46, 2425–2430 (1963)
Corrodi, H., Hillarp, N.-Å.: Fluoreszenzmethoden zur histochemischen Sichtbarmachung von Monoaminen. 2. Identifizierung des fluoreszierenden Produktes aus Dopamin und Formaldehyd. Helv. Chim. Acta 47, 911–918 (1964)
Costa, M., Gabella, G.: Adrenergic innervation of the alimentary canal. Z. Zellforsch. 122, 357–377 (1971)
Eränkö, O., Räisänen, L.: Demonstration of catecholamines in adrenergic nerve fibres by fixation in aqueous formaldehyde solution and fluorescence microscopy. J. Histochem. Cytochem. 14, 690–691 (1966)
Falck, B.: Observations on the possibilities of the cellular localization of monoamines by a fluorescence method. Acta Physiol. Scand. 56, Suppl. 197, I (1962)
Falck, B., Hillarp, N.-Å., Thieme, G., Torp, A.: Fluorescence of catecholamines and related compounds condensed with formaldehyde. J. Histochem. Cytochem. 10, 348–354 (1962)
Furness, J.B., Costa, M.: The adrenergic innervation of the gastrointestinal tract. Ergeb. Physiol. 69, 1–51 (1974)
Furness, J.B., Costa, M.: The use of glyoxylic acid for the fluorescence histochemical demonstration of peripheral stores of noradrenaline and 5-hydroxytryptamine in whole mounts. Histochemistry 41, 335–352 (1975)
Furness, J.B., Costa, M., Wilson, A.J.: Water-stable fluorophores, produced by reaction with aldehyde solutions, for the histochemical localization of catechol- and indolethylamines. Histochemistry 52, 159–170 (1977)
Furness, J.B., Malmfors, T.: Aspects of the arrangement of the adrenergic innervation in guinea-pigs as revealed by the fluorescence histochemical method applied to stretched, air-dried preparations. Histochemie 25, 297–309 (1971)
Hamberger, B., Norberg, K.A.: Histochemical demonstration of catecholamines in fresh frozen sections. J. Histochem. Cytochem. 12, 48–49 (1964)
Heene, R.: Histochemischer Nachweis von Katecholaminen und 5-Hydroxytryptamin im Kryostatschnitt. Histochemie 14, 324–327 (1968)
Håkansson, R., Owman, Ch., Sjöberg, N.-O., Sporrong, B.: Amine mechanisms in enterochromaffin and enterochromaffin-like cells of gastric mucosa in various mammals. Histochemie 21, 189–220 (1970)
Hökfelt, T., Ljungdahl, Å.: Modification of the Falck-Hillarp formaldehyde fluorescence method using the VibratomeR: simple, rapid and sensitive localization of catecholamines in sections of unfixed or formalin fixed brain tissue. Histochemie 29, 325–339 (1972)
Jacobowitz, D.: Histochemical studies of the autonomic innervation of the gut. J. Pharmacol. Exp. Ther. 149, 358–365 (1965)
Laties, A.M., Lund, R., Jacobowitz, D.: A simplified method for the histochemical localization of cardiac catecholamine-containing nerve fibres. J. Histochem. Cytochem. 15, 535–541 (1967)
la Torre, J.C. de, Surgeon, J.W.: A methodological approach to rapid and sensitive monoamine histofluorescence using a modified glyoxylic acid technique: the SPG method. Histochemistry 49, 81–93 (1976)
Lindvall, O., Björklund, A.: The glyoxylic acid fluorescence histochemical method: a detailed account of the methodology for the visualization of central catecholamine neurons. Histochemistry 39, 97–127 (1974)
Lindvall, O., Björklund, A.: Organization of catecholamine neurons in the rat cerebral nervous system. In: Handbook of Psychopharmacology, vol. 9. L.I. Iversen, S.D. Iversen and S.S. Snyder (eds.), pp. 139–231. New York, London: Plenum Press 1978
Lorén, I., Björklund, A., Falck, B., Lindvall, O.: An improved histofluorescence procedure for freeze-dried paraffin embedded tissue based on combined formaldehyde-glyoxylic acid perfusion with high magnesium content and acid pH. Histochemistry 49, 177–192 (1976)
Lorén, I., Björklund, A., Lindvall, O.: Magnesium ions in fluorescence histochemistry. Application to the cryostat and vibratome techniques. Histochemistry 52, 223–239 (1977a)
Lorén, I., Björklund, A., Falck, B., Lindvall, O.: The use of magnesium ions for sensitive visualization of catecholamines and serotonin in the CNS. Acta Physiol. Scand., Suppl. 452, 15–18 (1977b)
Lorén, I., Björklund, A., Falck, B., Lindvall, O.: The aluminum-formaldehyde (ALFA) histofluorescence method for improved visualization of catecholamines and indolamines. Application on the central nervous system. J. Neurosci. Meth., in press (1980)
Malmfors, T.: Studies on adrenergic nerves. Acta Physiol. Scand. 64, Suppl. 248, 1–93 (1965)
McLean, J.R., Burnstock, G.: Histochemical localization of catecholamines in the urinary bladder of the toad (Bufo marinus). J. Histochem. Cytochem. 15, 538–548 (1966)
Mellander, A., Ericson, L.E., Sundler, F., Ingbar, S.H.: Sympathetic innervation of the mouse thyroid and its significance in thyroid hormone secretion. Endocrinology 94, 959–966 (1974)
Mellander, A., Sundler, F., Westgren, U.: Sympathetic innervation of the thyroid: variation with species and with age. Endocrinology 96, 102–106 (1975)
Nelson, J.S., Wakefield, P.L.: The cellular localization of catecholamines in frozen-dried, cryostat sections of the brain and autonomic nervous system. J. Neuropath. Exp. Neurol. 27, 221–223 (1968)
Nielsen, K.C., Owman, Ch.: Difference in cardiac adrenergic innervation between hibernators and non-hibernating mammals. Acta Physiol. Scand., Suppl. 316 (1967)
Norberg, K.-A.: Transmitter histochemistry of the sympathetic adrenergic nervous system. Brain Res. 5, 125–170 (1967)
Nygren, L.-G.: On the visualization of central dopamine and noradrenaline nerve terminals in cryostat sections. Med. Biol. 54, 278–285 (1976)
Owman, Ch., Sjöberg, N.-O., Svensson, K.-G., Walles, B.: Autonomic nerves mediating contractility in the human Graafian follicle. J. Reprod. Fertil. 45, 553–556 (1975)
Placidi, G.F., Masouka, D.: Histochemical demonstration of fluorescent catecholamine terminal in cryostat sections of brain tissue. J. Histochem. Cytochem. 16, 491–492 (1968)
Sachs, Ch.: Noradrenaline uptake mechanisms in the mouse atrium. A biochemical and histochemical study. Acta Physiol. Scand., Suppl. 341, (1970)
Springgs, T., Lever, J., Rees, P., Graham, J.: Controlled formaldehyde-catecholamine condensation in cryostat sections to show adrenergic nerves by fluorescence. Stain Technol. 41, 323–327 (1966)
Stefenson, A., Owman, Ch., Sjöberg, N.-O., Walles, B.: Comparative study of the autonomic innervation of the mammalian ovary, with particular regard to the follicular system. Glyoxylic acid and cholinesterase histochemistry on vibratome sections. Submitted to Cell Tissue Res. (1979)
Waris, T., Partanen, S.: Demonstration of catecholamines in peripheral adrenergic nerves in stretch preparations with fluorescence induced by aqueous solution of glyoxylic acid. Histochemistry 41, 369–372 (1975)
Watson, J.S., Ellison, J.P.: Cryostat technique for central nervous system histofluorescence. Histochemistry 50, 119–127 (1976)
Watson, S.J., Barchas, J.D.: Catecholamine histofluorescence using cryostat sectioning and glyoxylic acid in unperfused frozen brain: a detailed description of the technique. Histochemical J. 9, 183–195 (1977)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ajelis, V., Björklund, A., Falck, B. et al. Application of the aluminum-formaldehyde (ALFA) histofluorescence method for demonstration of peripheral stores of catecholamines and indolamines in freeze-dried paraffin-embedded tissue, cryostat sections and whole-mounts. Histochemistry 65, 1–15 (1979). https://doi.org/10.1007/BF00496681
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00496681