JoVE Encyclopedia of Experiments
Neuroscience
0 views • 3:25 min • June 17th, 2025
Begin with an awake mouse positioned under a two-photon holographic microscope integrated with spatial light modulators or SLMs.
The mouse holds a pre-implanted head plate.
In the mouse brain, neurons express red-light-sensitive channels and green-fluorescent calcium indicators that bind to calcium ions.
To study neural activity and connectivity, set the imaging parameters.
Use a 920 nanometers laser to excite calcium-bound indicators within the neurons. Capture the neuron's fluorescence image with minimal photodamage.
Reset the imaging parameters and focus a near-infrared laser beam.
SLMs shape the laser beam into precise holographic patterns that allow selective focusing on target neurons at multiple points.
This stimulates light-sensitive channels, causes calcium ion influx, and enhances the target neurons' fluorescence.
The activated target neurons transmit signals to the connected neurons and increase the connected neuron's fluorescence.
Again, capture a two-photon image. An increased fluorescence in target and connected neurons confirms neuronal activity and connectivity.
After placing the mouse under the microscope and performing two-photon imaging, open the commercial imaging softwa
View the full transcript and gain access to thousands of scientific videos