An enzyme-linked immunosorbent assay for bromodeoxyuridine incorporation using fixed microcultures.
We report a quantitative method by which a single microculture can be examined for (i) cell morphology; (ii) cell number; (iii) DNA synthesis; and (iv) expression of cell antigens. This method first involves measuring by enzyme-linked immunosorbent assay (ELISA) the total bromodeoxyuridine (BrdU) incorporation into DNA by monolayer microcultures. The BrdU-ELISA measurement was followed by simultaneous immunostaining for BrdU-positive nuclei and for a cytoplasmic antigen. The method was applied to the measurement of mitogen-induced proliferation of rat sciatic nerve Schwann cell and cerebral astroglia microcultures. The ELISA measurement of BrdU incorporation compares favorably with measurements of tritiated thymidine incorporation and offers the additional advantages that the same microculture can subsequently be examined for cell number, for cell morphology, and for the percentage of cells having BrdU-labeled nuclei and other antigens.
Muir
D
D
Department of Biology, School of Medicine, University of California, San Diego, La Jolla 92093.
Varon
S
S
Manthorpe
M
M
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000276:immunology; D001253:Astrocytes / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000276:immunology / Q000378:metabolism*; D002452:Cell Count; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D003124:Colorimetry / Q000379:methods; D003470:Culture Media; D004247:DNA / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006041:Goats; D007118:Immunoassay; D051381:Rats; D012583:Schwann Cells / Q000378:metabolism / Q000648:ultrastructure; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D000906:Antibodies; D000941:Antigens; D003470:Culture Media; D014316:Tritium; D004247:DNA; D002772:Cholera Toxin; D001973:Bromodeoxyuridine; D013936:Thymidine