title: Stabilization media increases recovery in paucicellular cerebrospinal fluid specimens submitted for flow cytometry testing.
abstract: BACKGROUND
  Flow cytometric immunophenotpying (FCI) of cerebrospinal fluid (CSF) and other paucicellular fluids has been demonstrated to have increased sensitivity in detection of lymphoma and leukemia when compared to cytomorphology [(1) de Graaf et al., Cytometry Part B 2011, 80B:271-281; (2) Szamosi et al., CLSI Document H56-A-Body Fluid Analysis for Cellular Composition; Approved Guideline, Wayne, PA: Clinical and Laboratory Standards Institute, 2006; (3) Kraan et al., Flow Cytometric Immunophenotyping of Cerebrospinal Fluid. Current Protocols in Cytometry, Hoboken, NJ: Wiley, 2008]. However, low cellularity has been an historical problem with these samples. Several studies indicate that immediate addition of a stabilization media (e.g., RPMI with fetal calf serum (FCS)) to CSF improves the cell yield for FCI [(1) de Graaf et al.]. Such stabilization medias can, however, significantly increase cost.
  
  
  METHODS
  We compared FCI results in CSF stabilized with RPMI 1640 (without additional additives) to results obtained using non-stabilized CSF. Samples were processed according to published CLSI guidelines [(2) Szamosi et al.].
  
  
  RESULTS
  About 98/105 (93%) CSF specimens stabilized with RPMI had adequate numbers of viable cells (>100) for performing FCI. About 65/217 (30%) CSF specimens without stabilization had adequate numbers of viable cells for analysis (70% either quantity not sufficient (QNS) or specimen viability below analytical limits).
  
  
  CONCLUSIONS
  Utilizing RMPI without FCS as a stabilization media results in increased cell yield and improved FCI results. We have found FCS is not required to achieve high quality results in FCI of paucicellular CSF specimens.
authors.affiliation: Department of Pathology, Vanderbilt University Medical Center, Nashville, Tennessee.
authors.forename: B
authors.initials: B
authors.lastname: Greig
authors.forename: M
authors.initials: M
authors.lastname: Stetler-Stevenson
authors.forename: J
authors.initials: J
authors.lastname: Lea
mesh_terms: D002555:Cerebrospinal Fluid / Q000166:cytology* / Q000187:drug effects* / Q000276:immunology; D003470:Culture Media / Q000494:pharmacology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping
chemical_list: D003470:Culture Media
keywords: cerebrospinal fluid; leukemia; lymphoma; stabilization media