[Optimization of gold immunolabeling techniques on ultrathin slides obtained from samples fixed with glutaraldehyde and osmium tetroxide and enclosed in epoxy resin].
An immunogold labeling technique was carried out on plants infected with CMV, fixed with glutaraldehyde and osmium tetroxide and embedded in araldite CY 212. The effect of the type of support-film used, the resin and manipulations of the grids during immunogold steps, were studied and are discussed. The antigenic activity of virus was restored by treating the sections with sodium metaperiodate. The very high non-specific reactions observed with the support-film or with the resin were eliminated by adding powdered skimmed milk or non-purified albumin into the buffers. Purified bovine serum albumin (grade V) or chicken albumin (grade III to V) were inefficient in reducing this non-specific background.
Delecolle
B
B
INRA, Station de Pathologie Végétale, Domaine St-Maurice, Montfavet, France.
Womdim
R N
RN
Marchoux
G
G
D000956:Antigens, Viral / Q000032:analysis*; D002212:Capsicum; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004853:Epoxy Resins; D005976:Glutaral; D006652:Histological Techniques; D007202:Indicators and Reagents; D016253:Microscopy, Immunoelectron / Q000379:methods*; D009029:Mosaic Viruses / Q000648:ultrastructure*; D009993:Osmium Tetroxide; D010946:Plants, Medicinal; D010947:Plants, Toxic; D014026:Nicotiana
D000956:Antigens, Viral; D004853:Epoxy Resins; D007202:Indicators and Reagents; D009993:Osmium Tetroxide; D005976:Glutaral
Optimisation de la technique de l'immunomarquage à l'or sur sections ultrafines réalisées à partir d'échantillons fixés à la glutaraldéhyde et au tétroxyde d'osmium et inclus dans une époxy-résine.