Confocal Imaging of Cholesterol–Lipid Raft in Cells

Fluorescent Bodipy-labeled analogs of sphingolipids and cholesterol are specifically utilized to study the lipid microdomains of the cell membrane. Bodipy-labeled sphingomyelin and cholesterol can visualize the membrane lipid microdomains in living cells based on their fluorescence properties and help in examining the relationship between membrane domains at the cell surface (Marks et al., 2008). K562 cells were treated with vehicle control or cedrol for 20 h and collected by centrifugation at 2000 rpm for 3 min at 4°C. The pellet was resuspended in serum-free medium and then incubated with 20 μM of each Bodipy-labeled sphingomyelin and cholesterol (Invitrogen, United States) in serum-free medium for 30 min at 4°C. Cells were then washed with cold serum-free medium and observed under a confocal microscope.