Heller, JP;
Odii, T;
Zheng, K;
Rusakov, DA;
(2019)
Imaging tripartite synapses using super-resolution microscopy.
Methods
10.1016/j.ymeth.2019.05.024.
(In press).
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Abstract
Astroglia are vital facilitators of brain development, homeostasis, and metabolic support. In addition, they are also essential to the formation and regulation of synaptic circuits. Due to their extraordinary complex, at times nanoscopic, morphology, underlying cellular mechanisms have not been satisfactorily investigated or fully understood. Especially fine astrocytic processes that can be found in the vicinity of synapses poses difficulties to traditional imaging techniques. Here, we describe a 3D three-colour super-resolution microscopy approach to unravel the nanostructure of tripartite synapses. The method is based on the SMLM technique direct stochastic optical reconstruction microscopy (dSTORM) which uses conventional fluorophore-labelled antibodies. This approach enables reconstructing the nanoscale localisation of individual astrocytic glutamate transporter (GLT-1) molecules surrounding presynaptic (bassoon) and postsynaptic (Homer1) protein localisations in fixed mouse brain sections. However, the technique is readily adaptable to other types of targets and tissues.
| Type: | Article |
|---|---|
| Title: | Imaging tripartite synapses using super-resolution microscopy |
| Location: | United States |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.ymeth.2019.05.024 |
| Publisher version: | https://doi.org/10.1016/j.ymeth.2019.05.024 |
| Language: | English |
| Additional information: | This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
| Keywords: | Astrocytes, Immunohistochemistry, SMLM, Super-resolution microscopy, Tripartite synapses, dSTORM |
| UCL classification: | UCL 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 Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Clinical and Experimental Epilepsy |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10075773 |
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