A new method for preliminary screening of anticancer drugs by Hadamard transform microscopic fluorescence image analysis was proposed. Rat liver 2c hepatocyte was usedas the standard cell and acridine orange (AO) was employed to trace the interaction of drugand cellular DNA. The results for five anticancer drugs, vincristine, cyclophosphamide,mustine, cis-platin and mitomycin-C are in accordance with the mechanism of the drugs binding to cellular DNA and this indicates that this method is suitable for screening cell cyclenonspecific anticancer drugs.