| Treatment |
Step |
Cryoprotectant |
Expose time |
Temperature |
|
1 |
1 |
1 M Ac |
1.5 min |
20°C |
| 2 |
2 M Ac |
1.5 min |
20°C |
| 3 |
3 M Ac+1 M DMSO+2 M EG+2 M PG |
1 min |
20°C |
| 4 |
3 M Ac+3 M DMSO+4 M EG+4 M PG |
1 min |
20°C |
| 5 |
3 M Ac+5 M DMSO+5 M EG+6 M PG+15% Suc+15% H+5% PVP |
1 min |
0°C |
| 6 |
3 M Ac+5 M DMSO+5 M EG+6 M PG+20% Suc+20% H+10% PVP |
1 min |
0°C |
|
2 |
1 |
1 M Ac |
1.5 min |
20°C |
| 2 |
2 M Ac |
1.5 min |
20°C |
| 3 |
3 M Ac+1 M DMSO+2 M EG+1 M Gly |
1 min |
20°C |
| 4 |
3 M Ac+3 M DMSO+4 M EG+3 M Gly |
1 min |
20°C |
| 5 |
3 M Ac+5 M DMSO +5 M EG+5 M Gly +15% Suc+15% H+5% PVP |
1 min |
0°C |
| 6 |
3 M Ac+5 M DMSO+5 M EG+5 M Gly+20% Suc+20% H+10% PVP |
1 min |
0°C |
|
3 |
1 |
1 M Ac |
1.5 min |
20°C |
| 2 |
2 M Ac |
1.5 min |
20°C |
| 3 |
3 M Ac+1 M DMSO+2 M EG+2 M MeOH |
1 min |
20°C |
| 4 |
3 M Ac+3 M DMSO+4 M EG+4 M MeOH |
1 min |
20°C |
| 5 |
3 M Ac+5 M DMSO +5 M EG+6 M MeOH+15% Suc +15% H + 5% PVP |
1 min |
0°C |
| 6 |
3 M Ac+5 M DMSO+5 M EG+6 M MeOH
+20% Suc+20% H+10% PVP |
1 min |
0°C |
|
4 |
1 |
1 M Ac |
1.5 min |
20°C |
| 2 |
2 M Ac |
1.5 min |
20°C |
| 3 |
3 M Ac+2 M EG+2 M PG+1 M Gly |
1 min |
20°C |
| 4 |
3 M Ac+4 M EG+4 M PG+3 M Gly |
1 min |
20°C |
| 5 |
3 M Ac+5 M EG+6 M PG+5 M Gly+15%
Suc+15% H+5% PVP |
1 min |
0°C |
| 6 |
3 M Ac+5 M EG+6 M PG+5 M Gly+20%
Suc+20% H+10% PVP |
1 min |
0°C |
|
5 |
1 |
1 M Ac |
1.5 min |
20°C |
| 2 |
2 M Ac |
1.5 min |
20°C |
| 3 |
3 M Ac+2 M EG+2 M PG+2 M MeOH |
1 min |
20°C |
4
5
6 |
3 M Ac +4 M EG+4 M PG+4 M MeOH |
1 min |
20°C |
| 3 M Ac+5 M EG+6 M PG+6 M MeOH+15% Suc+15% H+5% PVP |
1 min |
0°C |
| 3 M Ac+5 M EG+6 M PG+6 M MeOH+20% Suc+20% H+10% PVP |
1 min |
0°C |
|
6 |
1 |
1 M Ac |
1.5 min |
20°C |
| 2 |
2 M Ac |
1.5 min |
20°C |
| 3 |
3 M Ac+2 M PG+1 M Gly+2 M MeOH |
1 min |
20°C |
| 4 |
3 M Ac+4 M PG+3 M Gly+4 M MeOH |
1 min |
20°C |
| 5 |
3 M Ac+6 M PG+5 M Gly+6 M MeOH+15% Suc+15% H+5% PVP |
1 min |
0°C |
| 6 |
3 M Ac+6 M PG+5M Gly+6 M MeOH+20% Suc+20% H+10% PVP |
1 min |
0°C |
