7. Single Quantum Dot Tracking of Membrane Receptors
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The dynamics of membrane proteins in living cells has become a major issue to understand important biological questions such
as chemotaxis, synaptic regulation, or signal transduction. The advent of semi-conductor quantum dots (QDs) has opened new
perspectives for the study of membrane properties because these new nanomaterials enable measurements at the single molecule
level with high signal-to-noise ratio. Probes used until now indeed encounter significant limitations: organic fluorophores
and fluorescent proteins rapidly photobleach, whereas gold particles and latex beads, although more stable, are bulky and
usually stain only one protein per experiment. In comparison, QDs are bright and photostable fluorescent probes with a size
on the order of 10 nm can be used with standard immunochemical methods. We present the experimental protocols and methods
of analysis which we used to investigate the dynamics of individual GABAA receptors in the axonal growth cone of spinal neurons in culture. Single QD tracking is nevertheless a general method, suitable
to study many transmembrane proteins.
Affiliation(s): (2) Laboratoire Kastler Brossel, Physics Department, École Normale Supérieure, Paris, France
(3) Laboratoire de Biologie Cellulaire de la Synapse, Biology Department, École Normale Supérieure, Paris, France
(3) Laboratoire de Biologie Cellulaire de la Synapse, Biology Department, École Normale Supérieure, Paris, France
Book Title: Quantum Dots: Applications in Biology
Series: Methods in Molecular Biology | Volume: 374 | Pub. Date: Feb-02-2007 | Page Range: 81-91 | DOI: 10.1385/1-59745-369-2:81
Subject: Biotechnology
Key Words: Quantum dots - single molecule tracking - axonal pathfinding - GABAA receptors - fluorescence microscopy - immunocytochemistry - chemotaxis
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