Vitrification and ultrarapid laser warming are crucial for the cryopreservation of animal embryos, oocytes, and other cells of medicinal, genetic, and agricultural value. In the present study, we focused on alignment and bonding techniques for a special cryojig that combines a jig tool and jig holder into one piece. This novel cryojig was used to obtain a high laser accuracy of 95% and a successful rewarming rate of 62%. The experimental results indicated that our refined device improved laser accuracy in the warming process after long-term cryo-storage through vitrification. We anticipate that our findings will lead to cryobanking applications that use vitrification and laser nanowarming to preserve cells and tissues from a wide range of species.

Vitrification and ultra-rapid laser warming technique is an important approach for cryopreservation of animal embryos and oocytes, as well as other cells with medicinal, genetic, and agricultural value. We believe that the long term cryo-storage after vitrification and for the following laser warming has only been achieved to date using our customized device which was first attempted to be developed in our study. In the present study, we focused on developing alignment and bonding techniques for special cryo-jig which were assembling jig tool and jig holder in one piece. This newly produced customized cryo-jig was demonstrated to have significantly high laser striking accuracy of 95% and a successful rewarming rate of 62%. This study was experimentally demonstrate an refined novel device for improvement of laser striking accuracy after long term cryo-storage using vitrification and laser warming technique. In addition, the customized device described herein was successfully applied to a biological sample with over a thousand coral larvae in long term cryo-storage. We anticipate that our core findings will provide further examples of cryobanking applications that use vitrification and nano-laser warming to help a wide range of cells and tissues from diverse species.