title: Imaging the lateral diffusion of membrane molecules with quantum dots.
abstract: 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.
authors.affiliation: INSERM U789, Biologie Cellulaire de la Synapse N&P, Ecole Normale Supérieure Paris, 46, Rue d'Ulm 75005 Paris, France.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Bannai
authors.forename: Sabine
authors.initials: S
authors.lastname: Lévi
authors.forename: Claude
authors.initials: C
authors.lastname: Schweizer
authors.forename: Maxime
authors.initials: M
authors.lastname: Dahan
authors.forename: Antoine
authors.initials: A
authors.lastname: Triller
mesh_terms: 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
chemical_list: D008563:Membrane Lipids; D008565:Membrane Proteins