Quantifying synapses: an immunocytochemistry-based assay to quantify synapse number.

Journal Article (Journal Article)

One of the most important goals in neuroscience is to understand the molecular cues that instruct early stages of synapse formation. As such it has become imperative to develop objective approaches to quantify changes in synaptic connectivity. Starting from sample fixation, this protocol details how to quantify synapse number both in dissociated neuronal culture and in brain sections using immunocytochemistry. Using compartment-specific antibodies, we label presynaptic terminals as well as sites of postsynaptic specialization. We define synapses as points of colocalization between the signals generated by these markers. The number of these colocalizations is quantified using a plug in Puncta Analyzer (written by Bary Wark, available upon request, c.eroglu@cellbio.duke.edu) under the ImageJ analysis software platform. The synapse assay described in this protocol can be applied to any neural tissue or culture preparation for which you have selective pre- and postsynaptic markers. This synapse assay is a valuable tool that can be widely utilized in the study of synaptic development.

Full Text

Duke Authors

Cited Authors

  • Ippolito, DM; Eroglu, C

Published Date

  • November 16, 2010

Published In

PubMed ID

  • 21113117

Pubmed Central ID

  • PMC3159596

Electronic International Standard Serial Number (EISSN)

  • 1940-087X

Digital Object Identifier (DOI)

  • 10.3791/2270


  • eng

Conference Location

  • United States