title: Flow cytometric immunophenotyping of cerebrospinal fluid.
abstract: Leptomeningeal disease is an important adverse complication occurring in patients with B and T cell lymphomas and acute leukemias of lymphoid and myeloid origin. Recent reports suggest that multiparameter flow cytometry immunophenotypic assessment of spinal fluid samples could improve the efficiency of detection of CNS involvement, due to its high specificity and greater sensitivity. However, spinal fluid samples are frequently paucicellular with a rapidly decreasing cell viability. Staining of spinal fluid therefore requires dedicated sample storage/transport, staining, and preparation protocols. The Basic Protocol in this unit outlines a consensus multiparameter (3- to 8-color) flow cytometry immunophenotypic protocol for the evaluation of CNS involvement of cerebrospinal fluid (CSF) samples by neoplastic cells. A Support Protocol describing the simultaneous assessment of surface and cytoplasmic antigens is also provided. Finally, in the Alternate Protocol, we describe a method to calculate absolute numbers of both normal and pathological cell subpopulations by adding counting beads to the assay.
authors.affiliation: Department of Medical Oncology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Jaco
authors.initials: J
authors.lastname: Kraan
authors.forename: Jan W
authors.initials: JW
authors.lastname: Gratama
authors.forename: Corinne
authors.initials: C
authors.lastname: Haioun
authors.forename: Alberto
authors.initials: A
authors.lastname: Orfao
authors.forename: Anne
authors.initials: A
authors.lastname: Plonquet
authors.forename: Anna
authors.initials: A
authors.lastname: Porwit
authors.forename: Sandra
authors.initials: S
authors.lastname: Quijano
authors.forename: Maryalice
authors.initials: M
authors.lastname: Stetler-Stevenson
authors.forename: Dolores
authors.initials: D
authors.lastname: Subira
authors.forename: Wyndham
authors.initials: W
authors.lastname: Wilson
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D002555:Cerebrospinal Fluid / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping / Q000379:methods; D007938:Leukemia / Q000150:complications / Q000473:pathology; D008223:Lymphoma / Q000150:complications / Q000473:pathology; D008577:Meningeal Neoplasms / Q000175:diagnosis
chemical_list: D000954:Antigens, Surface
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