{"id":96,"date":"2010-06-22T18:53:10","date_gmt":"2010-06-22T18:53:10","guid":{"rendered":"https:\/\/wordpress-dev.med.unc.edu\/mmrrc\/services\/unc-cryogenics\/sperm-thawing-protocol\/"},"modified":"2018-04-10T12:48:10","modified_gmt":"2018-04-10T16:48:10","slug":"sperm-thawing-protocol","status":"publish","type":"page","link":"https:\/\/www.med.unc.edu\/mmrrc\/services\/unc-cryogenics\/sperm-thawing-protocol\/","title":{"rendered":"Sperm Thawing Protocol"},"content":{"rendered":"

<\/p>\n

<\/p>\n

<\/p>\n

Samples must be stored in liquid nitrogen (or liquid nitrogen vapor) until ready to thaw<\/strong><\/h3>\n

Sperm Thawing<\/h4>\n
    \n
  1. Using forceps, grasp straw and hold in air for ~10 seconds then gradually lower into 37* water bath (10ul sperm fraction first, slowly lower until buffer fraction is in water) until ice disappears (~10 seconds).<\/li>\n
  2. Remove from water and gently wipe straw dry (straw may be fractured, but sample is OK)<\/li>\n
  3. Holding firmly, carefully cut off both sealed ends, beveling end with the 10ul sperm aliquot.<\/li>\n
  4. Transfer only the thawed sperm suspension directly into a 500ul IVF drop (do not let larger buffer column contact media).<\/li>\n
  5. Allow sperm to capacitate 40-60 minutes before adding oocytes.<\/li>\n
  6. Continue with IVF as usual.<\/li>\n<\/ol>\n

    Reference<\/h4>\n

    Conserving, Distributing and Managing Genetically Modified Mouse Lines by Sperm Cryopreservation G. Charles Ostermeier, Michael V. Wiles, Jane S. Farley, Robert A. Taft<\/p>\n","protected":false},"excerpt":{"rendered":"

    Samples must be stored in liquid nitrogen (or liquid nitrogen vapor) until ready to thaw Sperm Thawing Using forceps, grasp straw and hold in air for ~10 seconds then gradually lower into 37* water bath (10ul sperm fraction first, slowly lower until buffer fraction is in water) until ice disappears (~10 seconds). Remove from water … Read more<\/a><\/p>\n","protected":false},"author":1369,"featured_media":0,"parent":40,"menu_order":8,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"footnotes":"","_links_to":"","_links_to_target":""},"class_list":["post-96","page","type-page","status-publish","hentry","odd"],"acf":[],"yoast_head":"\nSperm Thawing Protocol - Mutant Mouse Resource and Research Centers at UNC<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.med.unc.edu\/mmrrc\/services\/unc-cryogenics\/sperm-thawing-protocol\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Sperm Thawing Protocol - Mutant Mouse Resource and Research Centers at UNC\" \/>\n<meta property=\"og:description\" content=\"Samples must be stored in liquid nitrogen (or liquid nitrogen vapor) until ready to thaw Sperm Thawing Using forceps, grasp straw and hold in air for ~10 seconds then gradually lower into 37* water bath (10ul sperm fraction first, slowly lower until buffer fraction is in water) until ice disappears (~10 seconds). 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