Flow cytometry for oligodendroglial analysis.

<p>Flow cytometry allows for rapid analysis of eight or more proteins expressed on individual cells. Cells are streamed through a narrow flow cell sequentially and interrogated individually, thus largely static organs such as the brain must be dissociated into individual cells prior to analysis. Cells are stained with a cocktail of antibodies, each conjugated to a unique fluorochrome. Because of distinct proteins changes that occur during the maturation of the oligodendrocyte, antibodies can be utilized to characterize cells throughout the oligodendrocyte lineage. Thousands of cells per second are streamed through the flow cytometer, excited by laser light, and emitted fluorescence is directed through filters to photomultiplier tubes specific for each fluorochrome. For subsequent analysis, single cells are distinguished from small debris using forward (FSC) and side (SSC) scatter of light. Live cells are distinguished from dead cells using a live/dead dye, and CNS resident cells are distinguished from residual hematopoietic cells using a CD45 antibody. The entire oligodendrocyte lineage, from early progenitor to mature oligodendrocyte, can be analyzed and quantified.