Imaging the lateral diffusion of membrane molecules with quantum dots.
This protocol describes a sensitive approach to tracking the motion of membrane molecules such as lipids and proteins with molecular resolution in live cells. This technique makes use of fluorescent semiconductor nanocrystals, quantum dots (QDs), as a probe to detect membrane molecules of interest. The photostability and brightness of QDs allow them to be tracked at a single particle level for longer periods than previous fluorophores, such as fluorescent proteins and organic dyes. QDs are bound to the extracellular part of the object to be followed, and their movements can be recorded with a fluorescence microscope equipped with a spectral lamp and a sensitive cooled charge-coupled device camera. The experimental procedure described for neurons below takes about 45 min. This technique is applicable to various cultured cells.
Bannai
Hiroko
H
INSERM U789, Biologie Cellulaire de la Synapse N&P, Ecole Normale Supérieure Paris, 46, Rue d'Ulm 75005 Paris, France.
Lévi
Sabine
S
Schweizer
Claude
C
Dahan
Maxime
M
Triller
Antoine
A
D002462:Cell Membrane / Q000378:metabolism*; D004058:Diffusion; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D015336:Molecular Probe Techniques; D045663:Quantum Dots
D008563:Membrane Lipids; D008565:Membrane Proteins