Improved immunocytochemical staining of carcinogen-DNA adducts by a capillary slot block system.
We developed an immunocytochemical protocol in which incubation occurs in a capillary slot instead of the conventional horizontal drop. Slots of constant width were formed by placing slides on top of each other with parafilm spacer layers in between. Cryostat or semi-thin plastic-embedded sections were cut from organs of carcinogen-treated experimental animals. Carcinogen-DNA adducts were visualized in the affected nuclei by a double peroxidase-antiperoxidase method using rabbit antisera specific for certain DNA adducts formed. The staining in capillary slot blocks offered better staining reproducibility than the conventional method. This is particularly important when the staining intensity must be quantified. In addition, handling of the blocks was substantially less laborious than the individual treatment of slides, making this protocol especially suitable for larger series of slides. Other applications for the capillary slot block protocol should be enzyme histochemistry and in situ hybridization.
Division of Chemical Carcinogenesis, The Netherlands Cancer Institute, Amsterdam.
E
E
Scherer
J
J
Van Benthem
D000818:Animals; D002273:Carcinogens / Q000032:analysis*; D002467:Cell Nucleus / Q000032:analysis; D004247:DNA / Q000032:analysis*; D006147:Guanine / Q000031:analogs & derivatives / Q000032:analysis; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000295:instrumentation / Q000379:methods*; D007728:Kupffer Cells / Q000648:ultrastructure; D008099:Liver / Q000032:analysis* / Q000187:drug effects; D051381:Rats; D013194:Staining and Labeling; D014021:Tissue Preservation / Q000379:methods
D002273:Carcinogens; D006147:Guanine; C008450:7-methylguanine; D004247:DNA