NONINVASIVE, REAL-TIME METHOD FOR THE EXAMINATION OF THYMIDINE UPTAKEEVENTS - APPLICATION OF THE METHOD TO V-79 CELL SYNCHRONY STUDIES

[C-14]Thymidine uptake into V-79 hamster lung fibroblasts has been suc cessfully demonstrated using a noninvasive, real-time method utilizing Cytostar-T scintillating microplates. These plates are standard forma t, tissue culture-treated, 96-well microplates with an integral scinti llating base. The microplates permit the culture and observation of ad herent cell monolayers. Biological activities of the cells can be stud ied by the provision of specific radiolabeled compounds, The biologica l activities of the adherent monolayer bring the specific radiolabel i nto proximity with the scintillating base and a scintillation signal i s thereby generated. [C-14]Thymidine incorporation on the microplates can be used to examine cell proliferation and cell cycle events, Using a combined mitotic shake-off/aphidicolin treatment to achieve synchro nization, the thymidine incorporation activities of V-79 cells have be en examined on Cytostar-T plates and correlated to traditional methods of determining incorporation. The method was further used to examine the effects of colcemid and olomoucine, both chemical inhibitors of ce ll proliferation, on synchronous populations of cells. The homogeneous detection format and the microplate nature of the method suggest a ro le for scintillating microplates in cell biology research and drug dis covery. (C) 1997 Academic Press.