Imaging Cytometry and the Diagnosis of Haematological Malignancies

Multiparameteric flow immunophenotyping has become the technique of choice for the analysis of haematological malignancies with diagnostic laboratories commonly analysing panels of 8–10 antibodies. Whilst this analysis allows detection of small changes in antigen expression for some haematological malignancies there is a need to correlate phenotype and genotype. Imaging flow cytometry combines high resolution digital images with quantitative information gained from standard flow. The imaging functionality allows the localisation of antigens to be determined (surface, cytoplasm, nuclear), a feature that cannot be achieved by standard flow. Specific populations can be selected for analysis based on fluorescent intensity, cell shape and size. Imaging flow has already been shown to have clinical applications in the analysis of haematological malignancies. Acute promyelocytic leukaemia (APML) can be diagnosed by detecting abnormal diffuse staining patterns of PML bodies in promyelocytes using the modulation feature for image analysis. Fluorescence in situ hybridisation (FISH) is a technique which uses probes to detect DNA sequences (e.g. 14q32/IGH in B lymphoid malignancies) and is clinically relevant for the diagnosis of a number of malignancies. Interpretation becomes difficult however where the number of genetically abnormal cells is low. Imaging cytometry enables FISH analysis of large numbers of cells identified by their phenotype, even when they only make up a subset of the sample. Whilst the place of imaging flow in the assessment of haematological malignancies is still undetermined, it has distinct potential for diagnostic assessment, prognostic stratification and providing information regarding therapeutic approach and monitoring of haemopoietic neoplasms. 

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