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Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number
Published on: November 16, 2010
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Quantifying molecular aggregation by super resolution microscopy within an excitatory synapse from mouse hippocampal
Shekhar Kedia1, Narendrakumar Ramanan1, Deepak Nair1
1Centre for Neuroscience, Indian Institute of Science, Bangalore 560012, India.
STAR Protocols
|May 3, 2021
Summary
This study introduces a super-resolution microscopy (SRM) method to analyze molecular organization in brain synapses. The technique reveals nanoscale differences within individual excitatory synapses, linking structure to function at high resolution.
Area of Science:
- Neuroscience
- Cell Biology
- Microscopy
Background:
- Super-resolution microscopy (SRM) is crucial for understanding molecular distribution at excitatory synapses.
- Existing methods may not fully capture nanoscale organization heterogeneity within neuronal subcompartments.
Purpose of the Study:
- To present a novel SRM paradigm for evaluating nanoscale organization heterogeneity between neuronal subcompartments.
- To identify morphological characteristics of nanodomains within functional zones of single excitatory synapses.
Main Methods:
- Utilized super-resolution microscopy (SRM) on mouse hippocampal neurons.
- Developed a protocol to identify and characterize nanodomains within synaptic subcompartments.
Main Results:
- Successfully identified morphological characteristics of nanodomains within functional zones of single excitatory synapses.
- Demonstrated the ability to analyze nanoscale organization heterogeneity between neuronal subcompartments.
Conclusions:
- The presented SRM paradigm enables correlation of structure and function at molecular resolution in single synapses.
- This protocol is adaptable for various immunocytochemical/histochemical samples and imaging paradigms.
