Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons

Luca Russo, Carmina Natale, Andrea Conz, Joe Kelk, Elena Restelli, Roberto Chiesa, Mario Salmona, Luana Fioriti, Luca Colnaghi

Research output: Contribution to journalArticlepeer-review


Synapses are the functional elements of neurons and their defects or losses are at the basis of several neurodegenerative and neurological disorders. Imaging studies are widely used to investigate their function and plasticity in physiological and pathological conditions. Because of their size and structure, localization studies of proteins require high-resolution imaging techniques. In this protocol, we describe a procedure to study in primary neurons the co-localization of target proteins with synaptic markers at a super-resolution level using structured illumination microscopy (SIM). SIM is a patterned-light illumination technique that doubles the spatial resolution of wide-field microscopy, reaching a detail of around 100 nm. The protocol indicates the required controls and settings for robust co-localization studies and an overview of the statistical methods to analyze the imaging data properly.

Original languageEnglish
JournalJ. Visualized Exp.
Issue number164
Publication statusE-pub ahead of print - Oct 31 2020


  • Biomarkers/metabolism
  • Imaging, Three-Dimensional
  • Microscopy/methods
  • Neurons/cytology
  • Protein Transport
  • Signal-To-Noise Ratio
  • Synapses/metabolism


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