A novel form of Total Internal Reflection Fluorescence Microscopy (LG-TIRFM) reveals different and independent lipid raft domains in living cells.
In the present study we have applied a novel form of Total Internal Reflection Fluorescence Microscopy (LG-TIRFM) in combination with fluorescently labeled cholera toxin to the study of lipid rafts dynamics in living cells. We demonstrate the usefulness of such approach by showing the dynamic formation/disaggregation of islands of cholera toxin on the surface of cells. Using multicolor LG-TIRFM with co-localization studies we show for the first time that two receptors previously identified as constituents of lipid rafts are found on different and independent "raft domains" on the cell plasma membrane. Furthermore, LG-TIRFM studies revealed limited association and dissociation of both domains overtime on different areas of the plasma membrane. The implications of different "raft domains" on cell physiology are discussed.
TIRF Technologies, 951 Aviation Parkway, Suite 700, Morrisville, NC 27560, USA.
Alexander
A
Asanov
Angélica
A
Zepeda
Luis
L
Vaca
D018958:CD55 Antigens / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D021962:Membrane Microdomains / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D020800:Receptor, Nerve Growth Factor / Q000378:metabolism
D018958:CD55 Antigens; D020800:Receptor, Nerve Growth Factor; D002772:Cholera Toxin