Simplified quantitative analysis of spinal cord cells from Theiler's virus-infected mice without the requirement for myelin debris removal

Flow cytometry is routinely used to analyze mononuclear cell populations in experimental CNS infections and autoimmune diseases in mice and other rodents. Previous analysis of mononuclear cell samples has involved centrifugation of single cell suspensions from minced CNS tissues on discontinuous gradients to separate cells from myelin debris: however, loss of cells that occurs on gradients necessitates pooling of CNS tissues. We have developed a method of analyzing cells in CNS tissues by flow cytometry without removing myelin debris, and applied this method to assess mononuclear cells and oligodendrocytes in SJL mice with Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease. This approach also allowed the determination of the TMEV burden in these cell populations.