Effect of tissue fixation on anti-bromodeoxyuridine immunohistochemistry.
We investigated the influence of various fixatives on monoclonal anti-BrdUrd antibody binding of BrdUrd-substituted DNA in tissue sections of routinely processed mouse small intestine after in vivo administration of BrdUrd. For denaturing fixatives such as ethanol or Carnoy's fluid, a standard denaturation protocol showed specific crypt cell labeling. With cross-linking agents such as formalin and glutaraldehyde, a remarkable increase in staining intensity was obtained after tissue digestion with pepsin before acid denaturation. The optimal pepsin concentration was determined for maximal immunoreactivity combined with acceptable morphology.
D000818:Animals; D000911:Antibodies, Monoclonal; D001973:Bromodeoxyuridine / Q000032:analysis* / Q000276:immunology; D003432:Cross-Linking Reagents; D004247:DNA / Q000032:analysis; D005404:Fixatives; D005557:Formaldehyde; D005629:Frozen Sections; D005976:Glutaral; D007124:Immunoenzyme Techniques; D007421:Intestine, Small / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D010434:Pepsin A; D011489:Protein Denaturation
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D005404:Fixatives; D005557:Formaldehyde; D004247:DNA; D010434:Pepsin A; D001973:Bromodeoxyuridine; D005976:Glutaral