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Related Experiment Video

Updated: Dec 28, 2025

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

Published on: November 14, 2016

Assessing Neuron-Astrocyte Spatial Interactions Using the Neuron-Astrocyte Proximity Assay.

Aina Badia-Soteras¹, J Christopher Octeau², Mark H G Verheijen¹

¹Department of Molecular and Cellular Neurobiology, Center for Neurogenomics and Cognitive Research, Amsterdam Neuroscience, VU University Amsterdam, Amsterdam, The Netherlands.

Current Protocols in Neuroscience

|February 19, 2020

Summary

This study details a Förster resonance energy transfer (FRET) method to visualize astrocyte process proximity to synapses in living brain tissue, aiding the study of neuron-astrocyte interactions.

Keywords:

astrocyte imaging neuron optical synapse

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Last Updated: Dec 28, 2025

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

Published on: November 14, 2016

Analyzing the Size, Shape, and Directionality of Networks of Coupled Astrocytes

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Published on: October 4, 2018

A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity

A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity

Published on: May 5, 2009

Area of Science:

Neuroscience
Cell Biology
Microscopy

Background:

Astrocytes form intimate contacts with synapses, influencing neuronal function.
Standard microscopy cannot resolve these nanoscale astrocyte-synapse interactions.
Understanding astrocyte proximity is key to their homeostatic and neuromodulatory roles.

Purpose of the Study:

To present a detailed protocol for analyzing astrocyte process proximity to synapses using Förster resonance energy transfer (FRET).
To enable the study of dynamic neuron-astrocyte interactions in live tissues.
To characterize astrocyte process proximity in the striatum.

Main Methods:

Development and application of a FRET-based approach for live-tissue imaging.
Utilizing adeno-associated virus microinjections and acute brain slice preparation.
Confocal microscopy and post hoc FRET quantification.

Main Results:

The FRET approach allows observation and tracking of static and dynamic astrocyte process proximity to synapses.
The protocol is compatible with standard imaging techniques.
Detailed protocols for cultured cells and acute brain slices are provided.

Conclusions:

The described FRET method provides a robust tool to study astrocyte-synapse interactions in vivo.
This technique facilitates a deeper understanding of astrocyte function in neural circuits.
The protocol enables characterization of astrocyte process proximity in living tissue.