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Multi-photon Intracellular Sodium Imaging Combined with UV-mediated Focal Uncaging of Glutamate in CA1 Pyramidal Neurons
Published on: October 8, 2014
16.0K
Combining Ca2+ imaging with -glutamate photorelease.
Marco Canepari1, Michel De Waard, David Ogden
1Inserm U836, Team 3, BP 170, Grenoble cedex 09, F-38042, France;
Cold Spring Harbor Protocols
|December 4, 2013
Summary
This study details methods for measuring optical calcium (Ca2+) signals evoked by glutamate photorelease. This technique isolates postsynaptic receptor activity, enabling analysis of distinct Ca2+ components.
Area of Science:
- Neuroscience
- Biophysics
- Optical Imaging
Background:
- Optical imaging of intracellular calcium (Ca2+) signals is crucial for understanding neuronal activity.
- Distinguishing Ca2+ influx through glutamate receptors from voltage-gated channels is challenging.
Purpose of the Study:
- To present simple configurations and methods for measuring optical Ca2+ signals.
- To enable the separation and analysis of different Ca2+ components by isolating postsynaptic glutamate receptor activation.
Main Methods:
- Photorelease of caged glutamate to activate postsynaptic receptors.
- Microscopy configurations optimized for illumination and tissue preservation.
- Two-photon illumination for simultaneous imaging and uncaging.
Main Results:
- Successful measurement of optical Ca2+ signals in response to glutamate photorelease.
- Demonstration of separating Ca2+ components originating from different sources.
- Methodology allows for analysis of postsynaptic receptor-mediated Ca2+ dynamics.
Conclusions:
- The described methods provide a robust approach for studying Ca2+ signaling.
- This technique facilitates the investigation of synaptic transmission and neuronal excitability.
- Optimized optical configurations and procedures enhance experimental efficiency and data quality.
