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A new assay for cytostatic drug testing is described which can be automated. Pleural effusions and ascites are cultured as such for one week. Cells of solid tumors are cultured in the patients own serum for the same time. The cells are then stained with the esterase and intracellular pH-indicator dye 1,4-diacetoxy-2,3-dicyano-benzene (ADB) to label vital cells. They are simultaneously stained with propidium iodide (PI) as an indicator for dead cells. Monosized fluorescent latex particles are added as concentration, volume and fluorescence standard. Inflammatory cells can be distinguished in the assay from tumor cells because of their small cell volume. The number of dead and surviving cells is counted by the flow cytometer and a therapeutic index is calculated as ratio between the surviving inflammatory to surviving tumor cells. An important feature of the assay is that the DNA-distribution of the dead cells (e.g. aneuploidy) as well as the functional state of the surviving tumor cells and inflammatory cells can be judged from intracellular esterase activity and intracellular pH.

An important intention of flow cytometric investigations is to obtain biochemical and biophysical information about cells which is suitable for automated tumor diagnosis. In this study, the ploidy status, the intracellular pH value, the intracellular esterase activity, and the cell volume of vital cells and the DNA and cell volume of dead cells were measured in cancerous tisse and normal lung tissue of 30 patients by flow cytometry. The cell samples were simultaneously stained with the pH and esterase indicator dye 1.4‐diacetoxy‐2,3‐dicyano‐benzene (ADB) and propidium iodide (PI). The flow cytometric measurements were performed in three‐parameter list mode. The data were evaluated on an AT‐compatible personal computer with the DIAGNOS1 program system for automated diagnosis of flow cytometric list mode data. Significant differences were found between normal and malignant tissue in DNA ploidy, in the intracellular esterase activity, in the cell, volume and in the percentage of inflammatory cells and parameters of necrosis. DNA‐aneuploidy was observed in 38% of the lung carcinomas. The simultaneous detection of DNA‐aneuploidy and tumor‐associated properties in a multifactorial analysis led to correct automatic tumor diagnosis in 85% of cases. DNA‐aneuploidy was found at a significant higher frequency in advanced tumors. Adenocarcinomas displayed DNA‐aneuploidy more often (80%) than squamous cell carcinomas (33%).

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