The Effect of Glass-cracking during Cooling in Liquid Nitrogen on Viability of Mint Shoot Tips
Cryopreservation using vitrification has been reported for several plant species. Shoot tips and vitrification solution were placed in semen straws and immersed in liquid nitrogen (LN). Cracking of the external glass occurred, but may be avoided by annealing slightly below the glass transition temperature before immersion. A varying percentage still cracked with some vitrification solutions. Rapid warming also can cause cracking. There is concern that cracking may reduce viability. Shoot tips from Mentha species were used to examine this problem. Glass cracking during either cooling or warming did not produce visible damage to shoot tips. Viability of shoot tips from tubes that cracked during cooling was not different from those that did not crack; however, shoot formation was slightly reduced. Cracking upon warming did not reduce viability nor shoot formation. Very slow warming reduced viability, but warming in either water or air (room temperature) gave higher levels of survival.