Alteration in DNA content is an early event in oral carcinogenesis. We have examined oral brush samples to detect non-diploid cells (NDC) using simultaneous morphological and cytogenetic analysis. The study included 8 oral squamous cell carcinomas (OSCC), 22 premalignant lesions (OPLs), and 25 control individuals. Slides stained with Giemsa followed by FISH using chromosome 2 centromeric DNA probe, were scanned and fluorescent signals were simultaneously analyzed in parallel with the morphology. The proportion of NDC increased with the severity of the diagnosis. In two control subjects, 1-1.5% of the examined cells were NDC. Over 2% NDC were present in all OSCC cases and in 11 of the OPLs, of which, in 8 the histologic diagnosis was either epithelial hyperplasia or mild dysplasia. A significant number of NDC had normal morphology when cytomorphology and FISH were compared. Two patients with OPLs developed OSCC these patients had a significant proportion of NDC. We suggest that the combined morphological and cytogenetic analysis of cells collected by a non-invasive brush sampling can enhance early detection of potentially malignant cells.