Skip to main content

Table 1 Freezing protocols

From: Comparison of conventional and automated freezing methods on PB2 rooster semen cryopreserved with glycerol and dimethylsulfoxide tris coconut-water extender

Protocols Freezing protocol 1 Freezing protocol 2 Freezing protocol 3 Freezing protocol 4
Cryo-vials type 0.25 mL straws 0.25 mL straws 0.25 mL straws 0.25 mL straws
Diluents Tris coconut water Tris coconut water Tris coconut water Tris coconut water
Cryoprotectant type Glycerol and DMSO Glycerol and DMSO Glycerol and DMSO Glycerol and DMSO
Cryoprotectant concentrations 8, 10, or 15% 8, 10, or 15% 8, 10, or 15% 8, 10, or 15%
Dilution rate 1:2 1:2 1:2 1:2
Equilibration time 4 h at 5 °C 4 h at 5 °C 4 h at 5 °C 4 h at 5 °C
Freezing rate − 3 °C/min until − 50 °C, − 50 °C/min to − 130 °C, finally− 196 °C 4 °C/min until − 50 °C, cool from − 50 to − 150 °C at a rate of 50 °C/min, finally − 196 °C l °C/min, to − 20 °C − 20 to − 80 °C at a rate of 20 °C/min, finally – 196 °C Vapor freezing at – 50 °C for 12 min, then immersed directly into liquid nitrogen at − 196 °C