JoVE Encyclopedia of Experiments
Neuroscience
0 views • 3:15 min • June 17th, 2025
Begin with a live postnatal mouse retinal explant mounted on a membrane filter and placed in an incubation chamber under a confocal microscope.
The retina is transfected to express a fluorescent protein in the membranes of starburst amacrine interneurons.
Place a pre-wetted sample weight onto the retina to stabilize it.
Fill the chamber with buffer.
Maintain optimal buffer flow and temperature in the chamber to preserve neuronal viability.
Position a water immersion objective into the imaging chamber.
Start epifluorescence imaging.
The laser light, directed through the objective, excites the fluorescent protein in the amacrine cells.
The emitted fluorescence is captured through the same objective, allowing visualization of the labeled amacrine cells.
Identify a brightly fluorescent cell for imaging.
Set the imaging parameters.
Capture images at multiple Z-planes to achieve a 3D visualization of the complete dendritic morphology.
Perform time-lapse imaging to track changes in dendritic morphology over time.
Assemble the live imaging incubation chamber and fill it with oxygenated aCSF. Turn on the pump and temperature controller, making sure that the temperature does not rise above 34 degrees Ce
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