Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2023, 167(2):121-130 | DOI: 10.5507/bp.2022.021

Analysis of cerebrospinal fluid cells by flow cytometry: Comparison to conventional cytology

David Zeman^{1, 2, 3, 4}, Kamila Revendova^{1, 4}, Radovan Bunganic⁴, Marketa Ryzi³, Petra Masarovicova³, Pavlina Kusnierova^{2, 3}, Vera Kotrlova^{2, 3}, Pavel Hradilek^{1, 4}, David Stejskal^{2, 3}, Vojtech Thon^{2, 5}

¹ Department of Clinical Neurosciences, Faculty of Medicine, University of Ostrava, Czech Republic

² Department of Biomedical Sciences, Faculty of Medicine, University of Ostrava, Czech Republic

³ Institute of Laboratory Medicine, University Hospital Ostrava, Czech Republic

⁴ Clinic of Neurology, University Hospital Ostrava, Czech Republic

⁵ RECETOX, Faculty of Science, Masaryk University, Brno, Czech Republic

Aims: This study compared the results obtained by basic immunophenotyping of cerebrospinal fluid (CSF) cells by flow cytometry (FC) to the results of conventional cytology and evaluated the possibility of detailed analyses of CSF B-cell subpopulations.

Methods: Samples from 42 patients were examined by conventional cytology (native and/or pre-centrifuged CSF) and FC. The results from 15 patients without evidence of organic neurological disease were used to estimate reference ranges.

Results: Pre-centrifugated CSF had significantly higher cell yield on the cytologic slide, but cell subpopulation percentages were altered; the percentage of lymphocytes was significantly higher and monocytes significantly lower compared to both native CSF slides and FC. The percentage of granulocytes was higher in FC compared to cytology. For leukocyte count, the following reference ranges were estimated for Fuchs-Rosenthal chamber (FR) counting and FC, respectively: leukocytes ≤4.7/μL and ≤2.5/μL, lymphocytes ≤4.1/μL and ≤1.8/μL, monocytes ≤1.2/μL and ≤0.9/μL, and granulocytes 0/μL and ≤0.2/μL. The following reference ranges were estimated for basic subpopulations: T-lymphocytes 84.1-100%, B lymphocytes 0.0-1.5%, NK cells 0.0-6.3%, NKT cells 0-9.5%, and CD3+CD4+/CD3+CD8+ 0.8-4.9. Using a volume of 1.2-2.4 mL, the number of B lymphocytes was too low (<20) in samples with ≤2.7 cells/μL in the FR.

Conclusions: Even normal CSF samples are amenable to basic mononuclear cell subpopulation analysis by FC. However, analysis of the B-cell subpopulations requires either a larger sample volume or selection of samples with ≥ 3 cells/μL.

Keywords: cerebrospinal fluid, cytology, flow cytometry, B lymphocytes

Received: December 14, 2021; Revised: April 6, 2022; Accepted: April 13, 2022; Prepublished online: May 4, 2022; Published: June 12, 2023  Show citation