SuperCLEM: an accessible correlative light and electron microscopy approach for investigation of neurons and glia in vitro.



Booth, Daniel G, Beckett, Alison J ORCID: 0000-0001-8377-325X, Prior, Ian A ORCID: 0000-0002-4055-5161 and Meijer, Dies
(2019) SuperCLEM: an accessible correlative light and electron microscopy approach for investigation of neurons and glia in vitro. Biology open.

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Abstract

The rapid evolution of super-resolution light microscopy has narrowed the gap between light and electron microscopy, allowing the imaging of molecules and cellular structures at high resolution within their normal cellular and tissue context. Multimodal imaging approaches such as correlative light electron microscopy (CLEM) combine these techniques to create a tool with unique imaging capacity. However, these approaches are typically reserved for specialists, and their application to the analysis of neural tissue is challenging. Here we present SuperCLEM, a relatively simple approach that combines super-resolution fluorescence light microscopy (FLM), 3D electron microscopy (3D-EM) and rendering into 3D models. We demonstrate our workflow using neuron-glia cultures from which we first acquire high-resolution fluorescent light images of myelinated axons. After resin embedding and re-identification of the region of interest, serially aligned EM sections are acquired and imaged using a serial block face scanning electron microscope (SBF-SEM). The FLM and 3D-EM data sets are then combined to render 3D models of the myelinated axons. Thus, the SuperCLEM imaging pipeline is a useful new tool for researchers pursuing similar questions in neuronal, as well as other complex tissue culture systems.

Item Type: Article
Depositing User: Symplectic Admin
Date Deposited: 04 Jun 2019 10:23
Last Modified: 20 Aug 2021 16:10
DOI: 10.1242/bio.042085
Open Access URL: http://10.0.4.218/bio.042085
URI: https://livrepository.liverpool.ac.uk/id/eprint/3044386