Improved method for immunoperoxidase detection of membrane antigens in frozen sections.
The visualisation of membrane antigens in frozen sections using metal-enhanced 3,3'diaminobenzidine (DAB) was investigated. Particular attention was paid to the degree of reaction enhancement, the disruptive effect on morphological detail, and the ease of the techniques. The best result was obtained using nickel-modified DAB at pH 6.0 to develop the peroxidase enzyme, with further enhancement in cobalt chloride at neutral pH. Silver methenamine enhancement is also possible but can give rise to non-specific or background staining.
Green
M A
MA
Department of Pathology, University of Newcastle upon Tyne.
Sviland
L
L
Malcolm
A J
AJ
Pearson
A D
AD
D015100:3,3'-Diaminobenzidine; D000954:Antigens, Surface / Q000032:analysis*; D003035:Cobalt; D003300:Copper; D005629:Frozen Sections / Q000379:methods; D006801:Humans; D007124:Immunoenzyme Techniques; D008867:Microtomy / Q000379:methods; D009532:Nickel; D014021:Tissue Preservation / Q000379:methods
D000954:Antigens, Surface; D015100:3,3'-Diaminobenzidine; D003035:Cobalt; D003300:Copper; D009532:Nickel