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Updated: Mar 28, 2026
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Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices
Published on: March 15, 2018
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Simultaneous Sodium and Calcium Imaging from Dendrites and Axons.
Kenichi Miyazaki1, William N Ross1
1Department of Physiology, New York Medical College, Valhalla, New York 10595; Marine Biological Laboratory, Woods Hole, Massachusetts 02543.
Eneuro
|January 6, 2016
Summary
Researchers developed a novel technique for simultaneous dynamic sodium and calcium imaging in neurons. This method enables high-speed, sensitive detection of intracellular ion changes, offering deeper insights into neural signaling pathways.
Area of Science:
- Neuroscience
- Cellular Physiology
- Biophysics
Background:
- Dynamic calcium imaging is crucial for neuroscience, revealing details on calcium channels, receptors, and intracellular calcium concentration ([Ca(2+)]i) dynamics.
- Dynamic sodium imaging offers analogous insights into sodium signaling but is less utilized.
- Simultaneous measurement of both intracellular calcium and sodium ([Na(+)]i) changes can provide more comprehensive data, especially for receptor signaling like AMPA and NMDA.
Purpose of the Study:
- To develop and validate a high-speed, sensitive technique for simultaneous dynamic imaging of intracellular sodium and calcium signals in single neurons.
- To overcome limitations of sequential imaging by capturing both signals concurrently in the same neuronal preparation.
- To enable a more complete understanding of neuronal signaling by correlating [Ca(2+)]i and [Na(+)]i dynamics.
Main Methods:
- Developed a custom optical setup with dichroic and emission filters for simultaneous detection of sodium and calcium indicators within a single neuron.
- Utilized high-intensity light-emitting diodes (LEDs) for alternate excitation of indicators, synchronized with a CCD camera operating at 500 Hz.
- Employed software to separate data streams, providing independent, high-resolution ([Ca(2+)]i) and ([Na(+)]i) signals from hippocampal brain slices.
Main Results:
- Achieved simultaneous, high-speed (500 Hz) imaging of intracellular sodium and calcium dynamics with submicron resolution.
- Successfully detected [Ca(2+)]i and [Na(+)]i changes associated with single action potentials in axons and synaptically evoked signals in dendrites.
- Demonstrated a good signal-to-noise ratio (S/N) for detecting localized physiological events.
Conclusions:
- The developed technique allows for simultaneous, high-resolution, and high-speed dynamic imaging of both intracellular sodium and calcium.
- This approach provides significant advantages over sequential imaging for studying neuronal signaling, particularly receptor-mediated events.
- The method enhances the ability to investigate the complex interplay between sodium and calcium dynamics in neural function.
