A method for the rapid analysis of neuronal proportions and neurite morphology in primary cultures.
This article provides basic guidelines for a rapid analysis of subpopulation proportions and neurite morphology in primary cultures. We describe, in E14 mesencephalic primary cultures, an immunohistochemical method for the simultaneous identification of multiple neuronal phenotypes and an estimation of the ratio of subpopulations. In addition, we describe the use of the Renaissance TSA-Direct kit (NEN, DuPont) to enhance the visualization of neurites when the antigen is in low abundance. Finally, a modified sholl analysis is used to rapidly and reliably estimate neurite number and length.
Isaacs
K R
KR
Laboratory of Clinical Science, NIMH, Bethesda, Maryland 20892, USA. Krystyna@helix.nih.gov
Hanbauer
I
I
Jacobowitz
D M
DM
D000818:Animals; D064032:Calbindin 2; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D005456:Fluorescent Dyes; D017408:Guidelines as Topic; D007150:Immunohistochemistry / Q000379:methods; D008636:Mesencephalon / Q000166:cytology*; D009419:Nerve Tissue Proteins / Q000032:analysis; D016501:Neurites / Q000648:ultrastructure*; D009474:Neurons / Q000166:cytology*; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D064030:S100 Calcium Binding Protein G / Q000032:analysis; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis
D064032:Calbindin 2; D005456:Fluorescent Dyes; D009419:Nerve Tissue Proteins; D011933:Reagent Kits, Diagnostic; D064030:S100 Calcium Binding Protein G; D014446:Tyrosine 3-Monooxygenase