Abstract
With correlative light and electron microscopy (CLEM), the ultrastructural cellular location of a biomolecule of interest can be determined using a combination of light microscopy (LM) and electron microscopy (EM). In many cases, the application of CLEM requires the use of markers that need to be attached to a biomolecule of interest to allow its identification and localization. Here, we review the potential of bioorthogonal chemistry to introduce such markers for CLEM.
Highlights
Correlative light and electron microscopy (CLEM) is an imaging technique that combines the virtues of light microscopy (LM) with those of electron microscopy (EM)
CLEM studies that involve fluorescence microscopy may benefit from fluorescent markers that can be attached to molecules of interest to allow their identification and localization
The examples and strategies discussed here highlight the power of bioorthogonal chemistry for the labelling of biomolecules in a cellular context
Summary
Correlative light and electron microscopy (CLEM) is an imaging technique that combines the virtues of light microscopy (LM) with those of electron microscopy (EM). After conversion to the nucleotide sugar CMPSiaNAz, SiaNAz is incorporated into various glycoconjugates by sialyltransferase enzymes With this approach, Saxon et al produced cells containing azide-tagged sialoglycans and visualized these using the Staudinger-Bertozzi reaction (Fig. 1b). The reaction procedure was reported to be simple, straightforward and non-toxic to E. coli cells (Fig. 1f) This variety in labelling strategies and chemical reactions highlights the versatility of the approach as it can be altered upon experimental settings. We anticipate that bioorthogonal chemistry will be a powerful and useful addition to the CLEM-labelling toolkit It would allow the imaging of non-protein biomolecules, and it precludes the need for genetic tagging and antibody labelling. We will highlight some of the inroads that have been made towards the CLEM imaging of bioorthogonal reactions
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