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Quantum dot conjugated nanobodies for multiplex imaging of protein dynamics at synapses

Modi, S; Higgs, NF; Sheehan, D; Griffin, LD; Kittler, JT; (2018) Quantum dot conjugated nanobodies for multiplex imaging of protein dynamics at synapses. Nanoscale , 10 (21) pp. 10241-10249. 10.1039/c7nr09130c. Green open access

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Abstract

Neurons communicate with each other through synapses, which show enrichment for specialized receptors. Although many studies have explored spatial enrichment and diffusion of these receptors in dissociated neurons using single particle tracking, much less is known about their dynamic properties at synapses in complex tissue like brain slices. Here we report the use of smaller and highly specific quantum dots conjugated with a recombinant single domain antibody fragment (VHH fragment) against green fluorescent protein to provide information on diffusion of adhesion molecules at the growth cone and neurotransmitter receptors at synapses. Our data reveals that QD-nanobodies can measure neurotransmitter receptor dynamics at both excitatory and inhibitory synapses in primary neuronal cultures as well as in ex vivo rat brain slices. We also demonstrate that this approach can be applied to tagging multiple proteins to simultaneously monitor their behavior. Thus, we provide a strategy for multiplex imaging of tagged membrane proteins to study their clustering, diffusion and transport both in vitro as well as in native tissue environments such as brain slices.

Type: Article
Title: Quantum dot conjugated nanobodies for multiplex imaging of protein dynamics at synapses
Location: England
Open access status: An open access version is available from UCL Discovery
DOI: 10.1039/c7nr09130c
Publisher version: http://doi.org/10.1039/c7nr09130c
Language: English
Additional information: This journal is © The Royal Society of Chemistry 2018. Open Access Article. Published on 17 May 2018. Downloaded on 01/06/2018 15:40:46. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Computer Science
URI: https://discovery.ucl.ac.uk/id/eprint/10049744
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