Rationale:

Flow cytometry is a method that allows simultaneous multiparametric analysis of the physical and/or chemical characteristics (for example, presence of specific cell surface and/or intracellular proteins) of single cells flowing through a fluorescence-activated cell sorter (FACS). In our program, we use this powerful technique to evaluate the cellular composition of various compartments of the immune system, such as thymus, Peyer's patches, spleen, lymph nodes, and peripheral blood as well as inflammatory infiltrates to various tissues in multiple disease models.

Methods:

The mice were euthanized by isoflurane overdose, their spleens were dissected and splenocyte suspensions were prepared. The splenocytes were stained for flow cytometry with anti-B220, anti-CD3, anti-CD4, anti-CD8, anti-pan-NK, anti-CD11b monoclonal antibodies. With respect to the flow cytometric analysis, 25,000 events (having the forward and side scatter properties of lymphocytes) were collected on a Becton Dickinson FACScan and analyzed using CellQuest software.