Surface Trafficking of Neurotransmitter Receptor: Comparison between Single-Molecule/Quantum Dot Strategies

Heterogeneity of AMPA receptor trafficking and molecular interactions revealed by superresolution analysis of live cell imaging

“…Interestingly, superresolution light optical microscopy techniques for in vivo data (1-3) have allowed monitoring a large number of molecular trajectories at the single molecule level and at nanometer resolution, that can potentially reveal unique cellular organizational features. In the recent years, various techniques based on empirical characterization have emerged to track receptors (4), and estimating the mean square displacement (MSD) along isolated trajectories allowed to differentiate between free and confined diffusion (5,6). In addition, although a large effort was dedicated to developing single molecule tracking algorithms (5,7,8), a general method for the analysis of the massive collection of data and for the extraction of quantitative local information is still lacking.…”

NTAㆍNi2+-Functionalized Quantum Dots for VAMP2 Labeling in Live Cells

“…The obvious answer for 'ideal' probes for future single molecule/particle tracking is to develop probes which are as small as possible, photo-resistant, and with very limited blinking. 5 We anticipated that NTA·Ni…”

“…4 However, streptavidin-coated QDs are quite large (20 -30 nm) and can impair the trafficking of labeled proteins in crowded cellular environments such as synapses. 5 The size of QDs increases when large proteins are used to functionalize the surfaces, e.g. streptavidin, which is 53 kDa, ~4 nm, or antibodies that are essential for labeling cell surface proteins.…”

Quantum Dots in Cell Biology

“…Second, QD-based SPT has been used to determine the diffusion characteristics of individual receptors as well as their subcellular localization over time (Bats et al 2007;Groc et al 2004;Groc et al 2007). It is important to note that diffusion Barroso measurements obtained using QD tracking methods have been generally consistent with those generated using fluorescence recovery photobleaching (FRAP) approaches (Bats et al 2007;Groc et al 2007). Third, individual or small assemblies of QD-ligand receptor complexes can be tracked for long periods of time in live cells (Gralle et al 2009;Groc et al 2004).…”