Abstract
A method for the cytochemical evaluation of the mechanism of action of potential metabolic inhibitors in cell culture, utilizing the rapid-scanning, high-resolution, automatically integrating biophysical instrumentation developed for cell population analyses is described. This method permits the in situ determination of the amount of DNA, RNA, and protein in individual cells in populations of heterogeneous cells, and the effects of inhibitory agents on these biosynthetic processes. The utility of the method has been demonstrated by the cytochemical evaluation of a series of inhibitory agents selected on the basis of what is known of their mechanisms of action with respect to inhibition of mitosis, synthesis of nucleic acids, or protein. The results of cytochemical analyses of populations of cells exposed to these agents agreed in general with what is known of their mechanisms of action from other studies. In addition to the advantages provided by a method based upon the evaluation of the effects of an inhibitory agent on single cells in heterogeneous populations, cytochemical analyses allow determination of the phase of the cell growth cycle in which inhibition occurs; information which is not readily obtained from other methods of assay. The advantages of such a method for the selection of candidate inhibitory agents for sequential or combination therapy of neoplastic disease, as well as detailed studies in cell physiology, are evident.
Published Version
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