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  1. Use large styrofoam box with liquid nitrogen high enough to submerge canes
  2. Take off the foam top of the storage dewar, identify the cane you need, and quickly and smoothly pull the cane out and drop it into the liquid nitrogen box.
  3. Transport that box near the hot water bath
  4. Hot water bath is adjusted to 25 degrees C.
  5. Identify the straw you want to thaw and grasp the center of the straw with metal tweezers under the surface of the liquid nitrogen (let the tweezers cool in liquid nitrogen before grabbing straw). In one second or less transfer the straw into the hot water bath. Keep the straw in the hot water bath for 15 seconds while constantly swirling.
  6. Remove the straw and dry with a Kimwipe and cut the sealed end of the straw.
  7. Place the newly cut end of the straw in a centrifuge tube on ice and carefully cut the opposite cotton end of the straw just below the cotton.
  8. Use straw filler syringe to empty straw into tube from the top.

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The “straw filler” is the syringe and pipette tip combination used in the CRYOKIT freezing video guide at timestamps 4:52-5:30. Depress the plunger of the syringe to dispense the sperm from the straw.

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