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NeuroScience Associates

Spinal Cord Preparation Post Fixation

 

If the spinal cord will not be removed soon after perfusion, then the dorsal parts of the vertebrae must be removed, as described below, so the cord is exposed to the fix solution (same as was used for the perfusion). Failure to do this can lead to a vacuole-like appearance in the tissue, especially the white matter. To prevent permanent curvature of spinal cord after perfusion fix the spinal column needs to be kept in a straight configuration for the first 24 hrs minimum. Significant curvature will have a deleterious effect on the final histologic product. After perfusion, the spinal column should be isolated from the rest of carcass (the head has already been decapitated).

  1. Using sturdy surgical scissors cut parallel to the spine separating the rib cage from the column.
  2. Cut the lower spinal column at a level even with the pelvis (upper hip joint).
  3. Remove as much as of the musculature as possible along the spinal column to minimize tissue mass.
  4. Using fine ronguers carefully expose the dorsal surface of the spinal cord to allow further penetration of fix.
  5. Using a suitable splint such as a tongue depressor, secure with sutures the spinal column to the splint and render the spinal column’s normal curvature straight. Take care that the sutures do not cut into the exposed spinal cord.
  6. Immerse the splint-spinal column in perfusion fix for at least 48 hours.
  7. Leave in fix at least overnight (for Amino CuAg, at least 12-16 hours).
  8. After appropriate time in fix solution, the spinal cord preparation can be switched to and shipped in buffer.

Removal of spinal cord immediately after perfusion:

After the cord is removed it will tend to assume the same curvature of the spinal column. To keep the cord straight, it can be tethered at both ends to a thin but sturdy piece of plastic, such as from X-ray film, as shown in this diagram. These ends will be damaged, and thus should not contain an area of interest.

SpinalCordSplint-web

Then follow steps 6-8.

Please contact our technical support team if you have any questions.

 

For more information:

Fixation Methods & Perfusion Solutions

Perfusion Protocol, Pump Calibration

Post-Fixation Buffer Solutions, Storage, Timing

Brain Extraction

Brain Hemisection

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