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Liquid nitrogen vapour freezing of mouse embryos

Summary. Mouse morulae were frozen rapidly to – 196°C in the presence of glycerol by a two-step procedure; the embryos were transferred directly from – 7°C after seeding into liquid nitrogen vapour at –170 to – 180°C and then into liquid nitrogen 10–15 min later. Suitable conditions for the survival of embryos frozen with liquid nitrogen vapour were found to be: 2 M-glycerol, 2 M-propylene glycol, 2 M-ethylene glycol; 5–30 min equilibration time at 0°C; 3–60 min holding time in liquid nitrogen vapour; dilution of glycerol with sucrose out of the frozen–thawed embryos; morula and early blastocyst stage embryos. Relatively high survival rates (69–74%) were obtained after rapid freezing by liquid nitrogen vapour. Morulae frozen in this fashion, cultured and transferred to recipients developed into normal young.