TY - JOUR
T1 - Improved contrast in histochemical detection of cytochrome oxidase
T2 - metallic ions protocol
AU - Divac, I.
AU - Mojsilovic-Petrovic, J.
AU - López-Figueroa, M. O.
AU - Petrovic-Minic, B.
AU - Møller, M.
N1 - Funding Information:
This work was supported by the Cultural Agreement between Denmark and Yugoslavia (J.M.-P.), the Danish Medical Research Council, the Danish Medical Association Research Fund, Fonden af 1870, Hassel-blad Foundation, Ib Henriksens Fond, Legat af 22. July 1959 and Michaelsens Fond. M.O.L.F. was supported by Ministerio de Asuntos Exteriores, Excmo. Cabildo de Tenerife and Fundacion Universitaria de Las Palmas and he gratefully acknowledges help from his family. Margaret Wong-Riley kindly provided a detailed description of her protocol. Povl Krogsgaard-Larsen was helpful in discussing the chemistry of the reaction. Samples of different Good’s buffers were kindly given to us by Svetid Bjiirklund, Bente Wis-chmann and Dorte Winther. Ilse Duun provided technical help and Margit L&green typed the manuscript. The initial phase of this work took place while I.D. was spending sabbatical leave at the Department of Neurology, UCLA School of Medicine, in the laboratory of R.C. Collins and E. Santori.
PY - 1995/2
Y1 - 1995/2
N2 - The standard current technique for demonstration of cytochrome oxidase (CyOx) provides low-contrast diaminobenzidine (DAB) polymer. In order to enhance the contrast with divalent metalic ions, we have screened a number of buffers and found that Hepes. Mops and cacodylate neither precipitate these ions nor inactivate CyOx in a concentration of 0.1 M. Staining thus obtained shows a broad range of gradations between black and white. With fresh tissue the resulting image is superior to that obtained with the brown DAB product, even if a recommended blue filter or printing on very hard paper are used. The technique is as simple as the one which is currently standard. Fixed tissue, cut either in a cryostat or vibratome, can be stained well when floating but not when mounted on slides. The stained floating tissue can be used for electron microscopy, but has no advantage over the standard method.
AB - The standard current technique for demonstration of cytochrome oxidase (CyOx) provides low-contrast diaminobenzidine (DAB) polymer. In order to enhance the contrast with divalent metalic ions, we have screened a number of buffers and found that Hepes. Mops and cacodylate neither precipitate these ions nor inactivate CyOx in a concentration of 0.1 M. Staining thus obtained shows a broad range of gradations between black and white. With fresh tissue the resulting image is superior to that obtained with the brown DAB product, even if a recommended blue filter or printing on very hard paper are used. The technique is as simple as the one which is currently standard. Fixed tissue, cut either in a cryostat or vibratome, can be stained well when floating but not when mounted on slides. The stained floating tissue can be used for electron microscopy, but has no advantage over the standard method.
KW - Diaminobenzidine
KW - Electron microscopy
KW - Nickel-ammonium sulfate
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U2 - 10.1016/0165-0270(94)00112-T
DO - 10.1016/0165-0270(94)00112-T
M3 - Article
C2 - 7752676
AN - SCOPUS:0028837182
SN - 0165-0270
VL - 56
SP - 105
EP - 113
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
IS - 2
ER -