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The detection of circulating tumour cells disseminated from solid tumours requires extremely sensitive methods. Molecular genetic methods, which are most sensitive, are not applicable to solid tumours because no tumour-specific genetic markers are available. Detection of disseminated tumour cells by immunocytochemistry is time-consuming, whereas fluorimetry is fast and quantitative. The laser scanning cytometer (LSC ® ) provides an automated microscopic procedure for screening up to 5×10 4  cells in suitable time. Using this system together with an enrichment procedure which allows up to ten thousand-fold enrichment, we have quantified minimal numbers of tumour cells. In a model system, breast cancer cell line cells diluted into peripheral blood mimicked seeding of tumour cells into the periphery. After staining with fluorochrome-conjugated anti-epithelial antibody, slides were screened for positive events directly or after enrichment with antibody-coated magnetic beads. One positive cell was unequivocally detectable in 10 4  cells and 50 out of 60 tumour cells were reliably recovered from a 20 ml blood volume, equal to 1–2 cells per 10 7 , after magnetic bead enrichment.

Detection and Quantification of Small Numbers of Circulating Tumour Cells in Peripheral Blood Using Laser Scanning Cytometer (LSC®)