LR White is preferable to Unicryl for immunogold detection of fixation-sensitive nuclear antigens.
The purpose of this study was to compare two electron microscopy embedding media - LR White and Unicryl - with regard to cell morphologyical and immunohistochemical preservation properties for the study of fixation-sensitive nuclear antigens. Human cervical carcinoma (HeLa) cells were fixed with 2% paraformaldehyde and 0.1% glutaraldehyde, and embedded in parallel in the two resins: LR White and Unicryl using; two different polymerization protocols were used for each resin. Preservation of fine nuclear structure was good after LR White and poor after Unicryl embedding. Immunogold labeling of Sm antigen was significantly stronger on LR White sections. Polymerization by UV light resulted in stronger and more specific labeling than heat polymerization. These results show that LR White is advantageous over Unicryl for the study of nuclear antigens requiring delicate aldehyde fixation.
Philimonenko
V V
VV
Institute of Experimental Medicine, Department of Cell Ultrastructure and Molecular Biology, Academy of Sciences of the Czech Republic, Vídenska 1083, 142 20 Prague 4 - Krc, Czech Republic.
Janácek
J
J
Hozák
P
P
D000180:Acrylic Resins; D034961:Antigens, Nuclear / Q000737:chemistry* / Q000302:isolation & purification; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D016253:Microscopy, Immunoelectron; D016610:Tissue Embedding; D016707:Tissue Fixation / Q000379:methods*; D014466:Ultraviolet Rays
D000180:Acrylic Resins; D034961:Antigens, Nuclear; D007202:Indicators and Reagents; C097557:unicryl; C048707:LR white