title: One tube with eight antibodies for 14-part bone marrow leukocyte differential using flow cytometry.
abstract: Bone marrow analysis by flow cytometry is part of the routine diagnosis of hematological disorders in medical laboratories. Differential leukocyte count and identification of abnormal cell subsets is currently performed through morphological examination on bone marrow smears by skilled cytologists. In this work, we propose a single 8-color tube for providing equivalent information, using flow cytometry.
  
  
  
  99 bone marrow samples were classified into 2 groups, (i) 51 samples, obtained from either healthy donors (n = 4) or patients with various diseases at diagnosis or during remission that did not present a hematological malignancy (n = 47), and (ii) 48 pathological samples with quantitative and/or qualitative abnormalities. A panel of eight antibodies-CD3-FITC/CD10-PE/CD38-PerCP-Cy5.5/CD19-PECy7/CD36-APC/CD16-APC-H7/CD34-BV421/CD45-V500-was tested to identify the main cell subsets at different stages of maturation using a FACSCanto-II analyzer.
  
  
  
  We first proposed a strategy of sequential gating leading to the identification of 14 leukocyte subsets, that is, erythroblasts, monocytes, B-lymphoid cells from hematogones to plasma-cells (5 subsets), T- and NK-cells, polymorphonuclear cells (neutrophils, eosinophils, and basophils), myeloblasts and other immature granular cells. This approach was validated by comparing flow cytometry and microscopic morphological examination, both in cases of normal and abnormal samples. Interestingly, cell identification, and numeration by flow cytometry was easy to perform and highly reproducible.
  
  
  
  A very simple, rapid, and reproducible flow cytometric approach, using a combination of eight antibodies allows determination of the cellular composition of bone marrow with high precision. © 2016 International Clinical Cytometry Society.
authors.affiliation: Université Grenoble-Alpes, Grenoble, 38000, France.|CNRS UMR 5309 and INSERM, U1209, Institut Albert Bonniot, Grenoble, 38706, France.|Department of Immunology CHU Grenoble, La Tronche, F-38700, France.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Jacob
authors.affiliation: Department of Immunology CHU Grenoble, La Tronche, F-38700, France.
authors.forename: Alice
authors.initials: A
authors.lastname: Souvignet
authors.affiliation: Department of Hematology, CHU Grenoble, La Tronche, F-38700, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Pont
authors.affiliation: Department of Hematology, CHU Saint Etienne, Saint Priest en Jarez, F-42270, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Solly
authors.affiliation: Department of Hematology, CHU Grenoble, La Tronche, F-38700, France.|Therex, TIMC-IMAG, CNRS Université Joseph Fourier, La Tronche, F-38700, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Mondet
authors.affiliation: Université Paris Diderot-CHU Saint Etienne, Saint Priest en Jarez, F-42270, France.
authors.forename: Sanae
authors.initials: S
authors.lastname: Kesr
authors.affiliation: Department of Immunology CHU Grenoble, La Tronche, F-38700, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Pernollet
authors.affiliation: Department of Immunology CHU Grenoble, La Tronche, F-38700, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Dumestre-Perard
authors.affiliation: Department of Hematology, CHU Saint Etienne, Saint Priest en Jarez, F-42270, France.
authors.forename: Lydia
authors.initials: L
authors.lastname: Campos
authors.affiliation: Department of Immunology CHU Grenoble, La Tronche, F-38700, France.
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Cesbron
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000906:Antibodies / Q000737:chemistry; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D001854:Bone Marrow Cells / Q000145:classification* / Q000276:immunology / Q000473:pathology; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression; D019337:Hematologic Neoplasms / Q000175:diagnosis* / Q000276:immunology / Q000473:pathology; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007223:Infant; D008214:Lymphocytes / Q000145:classification* / Q000276:immunology / Q000473:pathology; D008875:Middle Aged; D022423:Myeloid Cells / Q000145:classification* / Q000276:immunology / Q000473:pathology; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D005456:Fluorescent Dyes
keywords: bone marrow; flow-cytometry; leukocyte differential; standardization