Isolation of brain parenchymal lymphocytes for flow cytometric analysis. Application to acute viral encephalitis

David N. Irani, Diane E. Griffin

Research output: Contribution to journalArticlepeer-review

63 Scopus citations


A strategy for the isolation of mononuclear cells from the brain parenchyma of mice with ongoing central nervous system (CNS) inflammation has been developed in order to permit flow cytometric (FCM) analysis of these cell populations. Sindbis virus (SV) encephalitis in mice is characterized morphologically by an infiltration of mononuclear cells into both brain parenchyma and cerebrospinal fluid (CSF). Perfused brain tissue from infected animals is collected, homogenized, and subjected to a mild enzymatic digestion. A sedimentation at unit gravity is performed to remove any large particulate debris, and the remaining tissue is then centrifuged over a modified density gradient which separates intact cells from smaller tissue fragments. Cells collected directly from these gradients can be stained with monoclonal antibodies and analyzed by FCM without further manipulation. Data generated by this method correlates with previous studies of SV encephalitis using immunohistochemical analysis of brain tissue sections to quantify mononuclear cell types. This suggests that representative samples of the cellular infiltrate are obtained using this technique. The approach however, offers the possibility of more sophisticated and quantitative analyses of CNS inflammatory cells which is unobtainable by tissue section staining.

Original languageEnglish (US)
Pages (from-to)223-231
Number of pages9
JournalJournal of Immunological Methods
Issue number2
StatePublished - Jun 3 1991


  • Brain parenchyma
  • Flow cytometry
  • Lymphocyte isolation

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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