Single-cell analysis of lipid rafts in lymphocytes and in T cell-containing immunoconjugates

Within the plasma membranes of many different cell types, certain membrane lipids, including cholesterol and sphingolipids, form lateral assemblies surrounded by unsaturated glycerophospholipids. The concentration of such membrane lipids and associated proteins results in the formation of microdomains termed lipid rafts" (or glycolipid-enriched membranes or detergent-insoluble glycosphingolipid-enriched domains). Proteins that possess saturated acyl chains are generally associated with lipid rafts. Lipid rafts are believed to be involved in a number of cellular processes including cell activation. When material is limiting, raft-associated proteins may be identified on single cells using microscopy. This unit describes the application of this technique in an immunological example, examining the location and movement of signal transduction complexes in single T lymphocytes and in interactive conjugates between T cells and antigen-presenting cells (APCs).