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Materials

• Fixed brain tissue
• Tubes (one per tissue): conical tube (50 or 15 ml capacity), cryotube (2-5 ml capacity), or whatever size that's appropriate for the brain. The container should be made of plastic.
• Gauze (for whole perfusion-fixed brain)
• Fixative solution (same composition as used for fixation of brain tissue)
• Parafilm
• Cold pack (0-4°C): A small one should be sufficient. The purpose of this is to prevent drastic temperature fluctuations inside the box during shipping rather than keeping it at 4°C.
• Ziploc bag (or equivalent)
• Styrofoam box
• Packing material
• Packing tape

Procedure

1. Figure out a container that is suitable size for brain tissue being shipped:
   • Whole perfusion-fixed rat brain: 50-ml conical tube
   • Whole perfusion-fixed mouse brain: 15-ml conical tube is probably sufficient for a whole mouse brain (not tested)
   • Slabs of a larger brain (e.g., primates): plastic specimen jar of sufficient size with screw top
   • Fixed slab of brain tissue or acute slices: cryotube (2-5 ml capacity); tissue must fit without bending – if a cryotube is too small, use 15-ml conical tube.
2. Label the tube(s).
3. For a whole brain:
   1. Soak gauze in the fixative and loosely wrap the brain with it. Make sure that the wrapped brain fits snugly in the tube -- it should not move around but can slide in and out of the tube without extra force.
   2. Add ~20 ml of the fixative into a 50-ml conical tube. (~6 ml into a 15-ml tube)
   3. Place the gauze-wrapped brain into the tube.
   4. Fill the tube completely with the fixative.
   5. Tighten the cap and wrap around with a piece of Parafilm. Wipe clean the exterior of the tube.
   6. Place the tube into a Ziploc bag. Eliminate air before closing the bag.
4. For a slab or slices:
   1. Fill ~3/4 of a cryotube with the fixative.
   2. Place the fixed tissue into the tube without bending.
      • Minimize handling to keep the tissue from mechanical stress. If it is small enough, use a disposable transfer pipet (with its tip cut off) to place it into the tube.
   3. Fill the tube completely with the fixative.
   4. Tighten the cap.  Wipe clean the exterior of the tube.
   5. Place the cryotube into a 50-ml conical tube with padding (e.g., gauze or Kimwipe). Tighten the cap.
   6. Place the tube into a Ziploc bag. Eliminate air before closing the bag.
5. Place the Ziploc bag containing tube(s) and a cold pack into a Styrofoam box. Fill the box with packing material so that the tube(s) and cold pack wouldn't move around.
6. Close the box and seal with packing tape. Now the box is ready for overnight shipment.