A methodological approach to rapid and sensitive monoamine histofluorescence using a modified glyoxylic acid technique: The SPG method
- 283 Downloads
A modified approach of the glyoxylic acid (GA) condensation reaction for the visualization of biogenic amines in tissue is described.
Cryostat sections are used from brain or extracerebral tissue in dog, monkey, rat and mouse and exposed for 3 s to a room temperature solution containing sucrose-potassium phosphate-glyoxylic acid (SPG). The tissues are air dried and heated in an oven for 5 min. The complete processing time from fresh tissue to microscopic examination takes 18 min. Morphologically sharp and brightly fluorescent monoamine-containing neurons, pre-and terminal axons are seen against a dark parenchymal background without drug pre-treatment. The SPG method retains the high specific sensitivity for monoamines previously described in the original technique but is, in addition, more rapid and simple and is easily accessible as a research tool to investigators inexperienced in histofluorescence techniques.
KeywordsMonoamine Biogenic Amine Research Tool Condensation Reaction Temperature Solution
Unable to display preview. Download preview PDF.
- Axelsson, S., Bjorklund, A., Falck, B., Lindvall, O., Svensson, L.A.: Glyoxylic acid condensation: A new fluorescence method for the histochemical demonstration of biogenic monoamines. Acta physiol. scand. 89, 57–62 (1973)Google Scholar
- Battenberg, E.L.F., Bloom, F.E.: A rapid, simple and more sensitive method for the demonstration of central catecholamine neurons and axons by glyoxylic acid induced fluorescence. I specificity. Psychopharm. Comm. 1, 3–13 (1975)Google Scholar
- Dahlström, A., Fuxe, K.: Evidence for the existence of monoamine neurons in the central nervous system. I. Demonstration of monoamines in cell bodies of brain stem neurons. Acta physiol. scand. 64, Suppl. 232 (1964a)Google Scholar
- Dahlström, A., Fuxe, K.: A method for the demonstration of monoamine containing nerve fibers in the central nervous system. Acta physiol. scand. 60, 293–294 (1964b)Google Scholar
- Davis, J.N., Carlsson, A.: The effect of hypoxia on monoamine synthesis, levels and metabolism in rat brain. J. Neurochem. 21, 783–790 (1973)Google Scholar
- de la Torre, J.C.: Catecholamines in the human diencephalon:: A histochemical fluorescence study. Acta neuropath. 21, 165–168 (1972)Google Scholar
- de la Torre, J.C.: Dynamics of Brain Monoamines, pp. 27–37. New York: Plenum Press 1972Google Scholar
- de la Torre, J.C., Surgeon, J.W.: Brain and tissue monoamine histofluorescence visualization in 18 min. Fed. Proc. 35, 242 (1976).Google Scholar
- Falck, B.: Observations on the possibilities of the cellular localization of monoamines by a fluorescence method. Acta physol. scand. 56, Suppl. 197 (1962)Google Scholar
- 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)Google Scholar
- Falck, B., Owman, Ch.: A detailed methodological description of the fluorescence method for the cellular demonstration of biogenic monoamines. Acta Univ. Lund. 2, No. 7 (1965)Google Scholar
- Hokfelt, T., Fuxe, K., Johansson, O., Ljungdahl, A.: Pharmaco-histochemical evidence of the existence of dopamine nerve terminals in the limbic cortex. Europ. J. Pharmacol. 25, 108–112 (1974)Google Scholar
- Lindvall, O., Bjorklund, A.: The glyoxylic acid fluorescence histochemical method: A detailed account of the methodology for the visualization of central catecholamine neurons. Histochemistry 39, 97–127 (1974a)Google Scholar
- Lindvall, O., Bjorklund, A.: The organization of the ascending catecholamine neuron systems in the rat brain as revealed by the glyoxylic acid fluorescence method. Acta phsiol. scand. (Suppl. 412), 1–48 (1974b)Google Scholar
- Lindvall, O., Bjorklund, A., Hokfelt, T., Ljungdahl, Å.: Application of the glyoxylic acid method to Vibratome sections for improved visualization of central catecholamine neurons. Histochemie 35, 31–38 (1973)Google Scholar
- Ungerstedt, U.: Stereotaxic mapping of the monoamine pathways in the rat brain. Acta physiol. scand., Suppl. 367, 1–48 (1971)Google Scholar
- Yu, M.C., Bakay, L., Lee, J.C.: Effects of hypoxia nad hypercapnic hypoxia on the ultrastructure of central nervous synapses. Exp. Neurol. 40, 114–125 (1973)Google Scholar