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Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number
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Chapters
- 00:00Title
- 01:17Introduction
- 02:16Preparing and Fixing Dissociated Purified Rat Retinal Ganglion Cells
- 05:33Imaging
- 07:04Image Analysis and Co-localized Puncta Quantification
- 09:18Preparing and Fixing Brain Sections
- 12:39Imaging of Tissue Sections
- 13:34Representative Imaging and Synapse Assay Results
- 17:15Conclusion
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.
Tags
QuantifySynapsesImmunocytochemistry-based AssaySynapse NumberNeuroscienceMolecular CuesObjective ApproachesSynaptic ConnectivitySample FixationDissociated Neuronal CultureBrain SectionsImmunocytochemistryCompartment-specific AntibodiesPresynaptic TerminalsPostsynaptic SpecializationColocalizationSignalsMarkersPuncta AnalyzerImageJ Analysis Software PlatformSynapse AssayCulture Preparation
