ImageStream promyelocytic leukemia protein immunolocalization: In search of promyelocytic leukemia cells

Authors

  • Peppino Mirabelli,

    1. CEINGE-Biotecnologie Avanzate, Naples, Italy
    2. Dipartimento di Biochimica e Biotecnologie Mediche, Università Federico II, Naples, Italy
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    • Conflict of interest: The authors PM, GS, CP, FD, EM, RDN, and LDV have no competing interests. The authors TG, RK, VV and DB are shareholders and employees of Amnis Corporation.

  • Giulia Scalia,

    1. CEINGE-Biotecnologie Avanzate, Naples, Italy
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    • Conflict of interest: The authors PM, GS, CP, FD, EM, RDN, and LDV have no competing interests. The authors TG, RK, VV and DB are shareholders and employees of Amnis Corporation.

  • Caterina Pascariello,

    1. CEINGE-Biotecnologie Avanzate, Naples, Italy
    2. Dipartimento di Biochimica e Biotecnologie Mediche, Università Federico II, Naples, Italy
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    • Conflict of interest: The authors PM, GS, CP, FD, EM, RDN, and LDV have no competing interests. The authors TG, RK, VV and DB are shareholders and employees of Amnis Corporation.

  • Francesca D'Alessio,

    1. CEINGE-Biotecnologie Avanzate, Naples, Italy
    2. Dipartimento di Biochimica e Biotecnologie Mediche, Università Federico II, Naples, Italy
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  • Elisabetta Mariotti,

    1. CEINGE-Biotecnologie Avanzate, Naples, Italy
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    • Conflict of interest: The authors PM, GS, CP, FD, EM, RDN, and LDV have no competing interests. The authors TG, RK, VV and DB are shareholders and employees of Amnis Corporation.

  • Rosa Di Noto,

    Corresponding author
    1. CEINGE-Biotecnologie Avanzate, Naples, Italy
    2. Dipartimento di Biochimica e Biotecnologie Mediche, Università Federico II, Naples, Italy
    • CEINGE-Biotecnologie Avanzate, Via Gaetano Salvatore 486, 80145 Naples, Italy
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    • Conflict of interest: The authors PM, GS, CP, FD, EM, RDN, and LDV have no competing interests. The authors TG, RK, VV and DB are shareholders and employees of Amnis Corporation.

  • Thaddeus C. George,

    1. Amnis Corporation, Seattle, 2505 Third Avenue, Suite 210, Washington
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  • Raymond Kong,

    1. Amnis Corporation, Seattle, 2505 Third Avenue, Suite 210, Washington
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  • Vidya Venkatachalam,

    1. Amnis Corporation, Seattle, 2505 Third Avenue, Suite 210, Washington
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  • David Basiji,

    1. Amnis Corporation, Seattle, 2505 Third Avenue, Suite 210, Washington
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  • Luigi Del Vecchio

    1. CEINGE-Biotecnologie Avanzate, Naples, Italy
    2. Dipartimento di Biochimica e Biotecnologie Mediche, Università Federico II, Naples, Italy
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    • Conflict of interest: The authors PM, GS, CP, FD, EM, RDN, and LDV have no competing interests. The authors TG, RK, VV and DB are shareholders and employees of Amnis Corporation.


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. © 2012 International Society for Advancement of Cytometry

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