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JoVE Encyclopedia of Experiments
Encyclopedia of Experiments: Neuroscience

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A Technique for the Prolonged Incubation of Neuronal Tissue of the Retina

 

A Technique for the Prolonged Incubation of Neuronal Tissue of the Retina

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Transcript

Take an intact mouse eye in artificial cerebrospinal fluid, or aCSF, and incise to isolate the eyecup containing the retina.

Treat with an enzymatic solution to remove the inner limiting membrane, exposing the neuronal cells.

Immerse the tissue in an inhibition solution to stop digestion, and wash to remove excess solution.

Place the tissue in a specialized incubator at a low temperature to slow cellular metabolism, preserving the tissue.

This incubator comprises a main chamber with a mesh for mounting tissues. The chamber is filled with aCSF, and its temperature and pH are controlled to preserve tissue functionality.

To eliminate microbial contamination, the aCSF is pumped into another chamber and treated with ultraviolet radiation to kill any microbes.

The sterilized aCSF is then pumped back into the main chamber.

To isolate the retina, remove the eyecup, separate the retina from the surrounding tissue, and return it to the incubator. 

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