Almost forty years have passed since flow cytometry was developed in the late 1960s. It has been used in cell biology, particularly the two major fields of cell surface antigens and cellular DNA content. Its rapid analysis of the latter led to the detection of quantitative choromosomal abnormality, which would otherwise require much time, experience, and especially mitotic cells. It was adopted initially for haematological disorders, but its use expanded to solid tumours, and even fixed tissues for retrospective analysis. Although DNA aneuploidy was proven to be a prognostic factor for limited tumours, its clinical significance has not been established yet for most tumours. Inappropriate sampling and flow cytometry determining procedures might have generated confusing evaluations. The advantages of flow cytometric analysis of DNA aneuploidy include: 1) removal of the requirement for mitotic cells, 2) rapid determination (a rate of thousands per second), 3) qualitative determination (more than five thousand cells per sample), 4) labour-saving determination, and 5) objective determination by an instrument. Conversely, disadvantages might include: 1) impossibility of detecting qualitative chromosomal abnormality, 2) difficulty with detecting subtle quantitative abnormalities, and 3) impossibility of identifying relevant specific chromosomes. However, the advantages clearly outweigh the disadvantages. The Standardization Committee of the Japan Cytometry Society has recently finished creating the guidelines for flow cytometric analysis of DNA aneuploidy. These include sampling, providing free cell suspension, staining with fluorochromes, determining by flow cytometry, evaluating, and outlining the terminology of DNA aneuploidy. It is hoped that all researchers will adopt and comply with these guidelines whenever they analyse DNA aneuploidy with flow cytometry, and that they will find the appropriate clinical significance of DNA aneuploidy with its strict determination and definition.
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