Bioorthogonal labeling of live dissociated and organotypic slice culture neurons

Abstract Over the past couple decades, the explosion in the development of high-resolution and super-resolution microscopy techniques has led to the need for the development of new protein labeling techniques. Click-labeling via genetic code expansion (GCE) has received particular attention given its potential has the ultimately small labelling probe for proteins. Click-labeling via GCE offers a reliable and sterically minimally demanding capacity to label proteins, but its application in non dividing cells such as neurons remains poorly exploited due to its low efficiency. Here, we describe a simple, efficient and reproducible protocol that allows to fluorescently label transmembrane proteins in live neurons using click-labeling via GCE, both in dissociated culture and organotypic brain slices.