Double labeling of proliferating neurons with anti-BrdU and anti-NeuN: An improved immunohistochemical technique utilizing microwave irradiation

The authors describe a double labeling immunohistochemical technique to facilitate the study of neuronal proliferation and migration. Proliferating fetal neuroblasts were labeled with 5-bromo-2'-deoxyuridine (BrdU), and the labeled mature neurons were identified with a monoclonal anti-BrdU antibody. To differentiate neurons from glia, the tissue was also treated with monoclonal anti-NeuN, an antibody that detects a nuclear antigen unique to neurons. Novel aspects of this procedure are the utilization of methacarn fixation of tissues and the denaturation of BrdU-labeled DNA with microwave irradiation rather than with either acid or alkali denaturation treatments. This method produces strong immunoreactivity for both BrdU labeled neurons and neurons reacting with anti-NeuN, thereby facilitating the quantitative study of the development of cortical cytoarchitecture.

Duke Scholars

Author Sidney M. Gospe Jr. Pediatrics, Neurology