Advances in flow cytometry – how many colours do you need?

Abstract

Technological advances in flow cytometry include increasingly sophisticated instruments and an expanding range of fluoro-chromes. These advances are making it possible to detect an increasing number of markers simultaneously on the one cell. In clinical cytometry, the use of multiple markers has several advantages. For example, populations can be analysed more comprehensively and efficiently. There is improved sensitivity in the assessment of small populations, which is critical in the evaluation of minimal residual disease. When few cells are available in samples such as cerebrospinal fluid (CSF), more information can be obtained from a single tube. Disadvantages of the use of multiple markers include greater challenges with instrument compensation, and higher levels of training required for pathologists and laboratory staff. Developments in data analysis have lagged behind advances in instrumentation and fluorochromes. Implementation depends on a highly skilled laboratory scientist. Eight-colour systems are becoming increasingly widely used in clinical flow cytometry. For example, the EuroFlow Consortium is developing standardised antibody panels with suitable fluorochromes for the diagnosis, classification and monitoring of treatment of haematological malignancies. These protocols are based on the use of eight-colour flow cytometry.