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Updated: Jun 6, 2026
08:42
Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm
Published on: October 4, 2018
Multimodal optical imaging combining voltage-sensitive dye ElectroFluor630 with genetically encoded calcium,
Katarina D Milicevic1,2, Amal M Abdulkadir1, Kaio S Tom1
1UConn Health, School of Medicine, Department of Neuroscience, Farmington, Connecticut, United States.
Neurophotonics
|June 5, 2026
Summary
This study evaluates EF-630 voltage-sensitive dye compatibility with genetically encoded neuronal activity indicators (GENIs) for dual optical imaging. Paired recordings reveal kinetic differences influenced by indicator choice and cell type, crucial for interpreting neural population activity.
Area of Science:
- Neuroscience
- Optical Imaging
- Molecular Biology
Background:
- Understanding neural circuits requires simultaneous measurement of specific neuron activity and overall network responses.
- Genetically encoded neuronal activity indicators (GENIs) offer cell-type specificity, while voltage-sensitive dyes (VSDs) provide pan-neuronal activity.
- Far-red VSDs like EF-630 may enable dual-color imaging with green GENIs.
Purpose of the Study:
- To assess the compatibility of the far-red VSD EF-630 with various green fluorescent protein-based GENIs.
- To evaluate EF-630 as an internal reference for GENIs in dual optical imaging of neuronal populations.
- To establish performance benchmarks for paired VSD-GENI measurements.
Main Methods:
- Mouse brain slices expressing GENIs (GCaMP6f, iGluSnFR, ASAP2s, ASAP5-Kv) were stained with EF-630.
- Extracellular stimulation and dual-wavelength population optical imaging were performed.
- Kinetic properties (dynamics, temporal summation) of VSD and GENI signals were compared.
Main Results:
- Dual recordings successfully captured population signals from EF-630 and all tested GENIs.
- Voltage indicators ASAP2s and ASAP5-Kv showed faster dynamics and less temporal summation than VSD signals.
- Observed kinetic differences were influenced by cell-type-specific expression of GENIs.
Conclusions:
- The choice of indicator pair and targeted cell type significantly impacts the interpretation of neural population activity.
- This study provides the first systematic characterization of paired VSD-GENI measurements.
- Practical considerations and performance benchmarks are established for dual optical imaging in neuroscience research.
