Protocol for studying GABA(A) receptor subsynaptic domains in rat hippocampal neurons using single-molecule localization microscopy.

The study of protein organization at synapses and their rearrangement to maintain efficient synaptic transmission is made possible by single-molecule super-resolution imaging. Here, we present a protocol for studying the organization of GABA(A)R subdomains at inhibitory synapses. We describe steps for cell fixation, immunolabeling, super-resolution acquisition, and data analysis using a MATLAB open-source algorithm. This protocol has potential applications in the study of membrane molecules at synapses, as well as in other cellular compartments. For complete details on the use and execution of this protocol, please refer to Merlaud et al.(1).