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Thawing sperm

This protocol may be used to thaw sperm that has been frozen in straws or cryovials.

Category
Protocol
Record History
Added on June 21, 2010 at 1:48 PM by Lindner, Jill
Modified on August 6, 2010 at 3:20 PM by Skelton, Jennifer
Shared with (contributions)
TMESCSR: Protocols

Reagents, supplies, and equipment:

  • paper clip plunger
  • IVF dish
  • scissors
  • 100% ethanol

Thawing sperm in straws:

Procedure:

  1. Remove straw from LN2 storage container.
  2. Immerse straw in room temperature water bath until all ice crystals are melted (approximately 2 minutes).
  3. Wipe off straw with ethanol.
  4. Cut off sealed end and cut at the center of the cotton plug.
  5. Using plunger, expel sperm fraction into dish of IVF media.

 

Thawing Sperm in Vials:

Procedure:

  1. Remove cryo tubes from LN2 storage and place on the bench at room temperature until thawed, usually 12 - 15 minutes.
  2. Using wide bore pipette, remove quantity of sperm needed for experiment.