UNIT 6.25 Flow Cytometric Immunophenotyping of Cerebrospinal Fluid

  1. Jaco Kraan1,
  2. Jan W. Gratama1,
  3. Corinne Haioun2,
  4. Alberto Orfao3,
  5. Anne Plonquet4,
  6. Anna Porwit5,
  7. Sandra Quijano3,
  8. Maryalice Stetler-Stevenson6,
  9. Dolores Subira7,
  10. Wyndham Wilson8

Published Online: 1 JUL 2008

DOI: 10.1002/0471142956.cy0625s45

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Kraan, J., Gratama, J. W., Haioun, C., Orfao, A., Plonquet, A., Porwit, A., Quijano, S., Stetler-Stevenson, M., Subira, D. and Wilson, W. 2008. Flow Cytometric Immunophenotyping of Cerebrospinal Fluid. Current Protocols in Cytometry. 45:6.25:6.25.1–6.25.16.

Author Information

  1. 1

    Department of Medical Oncology, Erasmus Medical Center, Rotterdam, The Netherlands

  2. 2

    Service d'Hématologie Clinique, Hospital Henri Mondor, Créteil, France

  3. 3

    CSIC, University of Salamanca, Salamanca, Spain

  4. 4

    Immunologie Biologique, Hospital Henri Mondor, Créteil, France

  5. 5

    Department of Pathology, Karolinska University Hospital, Stockholm, Sweden

  6. 6

    Flow Cytometry Unit, National Institutes of Health, Bethesda, Maryland

  7. 7

    Department of Hematology, Jiménez Díaz Foundation, Madrid, Spain

  8. 8

    Center for Cancer Research, National Institutes of Health, Bethesda, Maryland

Publication History

  1. Published Online: 1 JUL 2008
  2. Published Print: JUL 2008


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. Curr. Protocol. Cytom. 45:6.25.1-6.25.16. © 2008 by John Wiley & Sons, Inc.


  • CSF;
  • cerebrospinal fluid;
  • diagnosis;
  • flow cytometry;
  • immunophenotyping;
  • leukemia;
  • lymphoma