ImageStream promyelocytic leukemia protein immunolocalization: in search of promyelocytic leukemia cells |
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Authors: | Mirabelli Peppino Scalia Giulia Pascariello Caterina D'Alessio Francesca Mariotti Elisabetta Di Noto Rosa George Thaddeus C Kong Raymond Venkatachalam Vidya Basiji David Del Vecchio Luigi |
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Institution: | CEINGE-Biotecnologie Avanzate, Naples, Italy. |
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Abstract: | Acute promyelocytic leukemia (APL) is a hematological emergency in which a rapid diagnosis is essential for early administration of appropriate therapy, including all-trans retinoic acid before the onset of fatal coagulopathy. Currently, the following methodologies are widely used for rapid initial diagnosis of APL: 1) identification of hypergranular leukemic promyelocytes by using classical morphology; 2) identification of cells with diffuse promyelocytic leukemia (PML) protein distribution by immunofluorescence microscopy; 3) evidence of aberrant promyelocyte surface immunophenotype by conventional flow cytometry (FCM). Here, we show a method for immunofluorescent detection of PML localization using ImageStream FCM. This technique provides objective per-cell quantitative image analysis for statistically large sample sizes, enabling precise and operator-independent PML pattern recognition even in electronic and real dilution experiments up to 10% of APL cellular presence. Therefore, we evidence that this method could be helpful for rapid and objective initial diagnosis and the prompt initiation of APL treatment. |
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