A Fresh Mount Method for Cytochrome Oxidase Histochemistry

Hippocampal and caudate metabolic activity associated with different navigational strategies.

“…Sections from the brains were used to perform CO histochemistry. We used a modified version of the method originally described by Wong-Riley (1979), which was based on the method developed by Sukekawa (1991). In brief, slides were lightly fixed for 5 min with a 1.5% glutaraldehyde solution, rinsed 3 times in phosphate buffer, and incubated at 37 °C for 2 hr in the dark and with continuous stirring in a solution that contained 50 mg 3,3Ј-diaminobenzidine, 15-mg cytochrome c (Sigma Chemical, St. Louis, Missouri), 4 g sucrose per 100 ml phosphate buffer (pH 7.4; 0.1 M).…”

Cytochrome Oxidase Activity of the Suprachiasmatic Nucleus and Pineal Gland in Rats with Portacaval Shunt

“…They were sectioned in a cryostat (Jung-Reichert, Frigocut-250, Heidelberg, Germany) at Ϫ20°C, and 20m-thick sections were obtained. The sections were histochemically processed to reveal CO by diaminobenzidine according to a modification of the Wong-Riley method (75) by Sukekawa (68). Alternate sections were Nissl stained (with cresyl violet), in order to locate the SCN (Fig.…”

Dopaminergic Control of Kynurenate Levels and N-Methyl-<i>D</i>-Aspartate Toxicity in the Developing Rat Striatum

“…Cryostat sections (30 Ìm) were cut coronally through the entire striatum, and one section out of four was collected on polylysinated slides and kept at -20°C. The sections were stained using a metal-enhanced cytochrome oxidase method [Silverman and Tootell, 1987;Sukekawa, 1991], replacing cobalt chloride with nickel ammonium sulfate. For this purpose, the sections were warmed to room temperature and incubated for 75 min in 100 ml of 0.1 M Hepes buffer, pH 7.5, containing 22.4 mg cytochrome c, 115 mg diaminobenzidine, 4.5 g sucrose and 12.5 ml of a 1% nickel ammonium sulfate solution.…”