Table 1 Methods and CPA concentrations used to cryopreserve P. lividus eggs and obtain SEM and TEM images.
From: Ultrastructural examination of cryodamage in Paracentrotus lividus eggs during cryopreservation
Cryopreservation method | CPA concentration | Cooling rates | Thawing rates |
---|---|---|---|
Slow cooling | 0.5 M, 1 M, 1.5 M, 2 M, 2.5 M, 3 M DMSO 1.5 M DMSO + 0.04 M TRE1.5 M DMSO + 0.75 M PVP 1.5 M DMSO + 0.2 M SUC 1.5 M DMSO + 1% BSA | 1 °C/min11 | 280 °C/min water bath at 25 °C61 |
Vitrification by contact | NONE 0.5 M, 1.5 M, 3 M MeSo2 0.5 M, 1.5 M, 3 M EG | CVM™ kit 10,000 °C/min62 | N/A |
Droplet vitrification | NONE 0.5 M, 1.5 M, 3 M DMSO 0.5 M, 1.5 M, 3 M EG | 3 and 5 mm droplets, 1320 and 960 °C/min respectively60 | 200 °C/min water bath at 37–40 °C63a |
Straw directly into LN2 | NONE 0.5 M, 1.5 M, 3 M DMSO 0.5 M, 1.5 M, 3 M EG | 1827 °C/min50 | 2950 °C/min water bath at 25 °C64 |