Catecholamine histofluorescence using cryostat sectioning and glyoxylic acid in unperfused frozen brain: a detailed description of the technique.
This paper presents a technique for glyoxylic acid-induced monoamine histofluorescence in the central nervous system. Unperfused rat brains are sectioned in a cryostat, immersed in 2% glyoxylic acid solution, warm-air dried and gassed at 100 degrees C. Intense, well-localized catecholamine fluorescence is produced and all known catecholamine-containing structures are demonstrated. The fluorescence obtained by this method was evaluated by a variety of agents and was shown to be catecholaminergic in origin. In contrast to the Vibratome-glyoxylic acid technique, this procedure reliably produces thin, whole-brain sections of even thickness and allows protracted use of the tissue block. Because unperfused tissue is used, the technique can be applied to a broad variety of material, such as post-mortem tissue or invertebrate preparations. Alternate sections can be prepared for a variety of techniques requiring unperfused tissue (e.g., enzymatic localization, chemical assay, anatomical techniques). The reasons for choosing each of the parameters in the technique are discussed.
D000818:Animals; D001369:Axons / Q000032:analysis; D001923:Brain Chemistry / Q000187:drug effects; D004298:Dopamine / Q000032:analysis*; D005453:Fluorescence; D005629:Frozen Sections; D006038:Glyoxylates; D006651:Histocytochemistry / Q000379:methods*; D007980:Levodopa / Q000494:pharmacology; D008297:Male; D009638:Norepinephrine / Q000032:analysis*; D051381:Rats; D012110:Reserpine / Q000494:pharmacology; D012701:Serotonin / Q000032:analysis*; D014364:Tryptophan / Q000494:pharmacology
D006038:Glyoxylates; D012701:Serotonin; D007980:Levodopa; D012110:Reserpine; D014364:Tryptophan; D004298:Dopamine; D009638:Norepinephrine