title: ImageStream promyelocytic leukemia protein immunolocalization: in search of promyelocytic leukemia cells.
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.
authors.affiliation: CEINGE-Biotecnologie Avanzate, Naples, Italy.
authors.forename: Peppino
authors.initials: P
authors.lastname: Mirabelli
authors.forename: Giulia
authors.initials: G
authors.lastname: Scalia
authors.forename: Caterina
authors.initials: C
authors.lastname: Pascariello
authors.forename: Francesca
authors.initials: F
authors.lastname: D'Alessio
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Mariotti
authors.forename: Rosa
authors.initials: R
authors.lastname: Di Noto
authors.forename: Thaddeus C
authors.initials: TC
authors.lastname: George
authors.forename: Raymond
authors.initials: R
authors.lastname: Kong
authors.forename: Vidya
authors.initials: V
authors.lastname: Venkatachalam
authors.forename: David
authors.initials: D
authors.lastname: Basiji
authors.forename: Luigi
authors.initials: L
authors.lastname: Del Vecchio
mesh_terms: D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods*; D042381:Granulocyte Precursor Cells / Q000502:physiology*; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D015473:Leukemia, Promyelocytic, Acute / Q000175:diagnosis*; D009363:Neoplasm Proteins / Q000032:analysis / Q000737:chemistry; D009687:Nuclear Proteins / Q000032:analysis* / Q000737:chemistry; D000071156:Promyelocytic Leukemia Protein; D013194:Staining and Labeling / Q000379:methods; D016707:Tissue Fixation / Q000379:methods; D014157:Transcription Factors / Q000032:analysis* / Q000737:chemistry; D025521:Tumor Suppressor Proteins / Q000032:analysis* / Q000737:chemistry
chemical_list: D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human