Novel Long-Term Cryo-Storage Using Vitrification and Laser Warming Techniques

Refined novel techniques for long term cryo-storage using vitrification and laser warming

“…The major function of the custom-made work holding cryo-jig is to guide the tool, whereas the fixture is a tool that securely and firmly keeps the work in place to complete a certain operation, and to adjust the manufacturing process in time to increase the laser striking accuracy and consistent vitrification rate, and assure high and successful laser rewarming. This newly produced customized cryo-jig was demonstrated to have significantly higher laser striking accuracy of more than 90% and a successful rewarming rate of 62% in comparison to that presented in our prior study of 76% and a rewarming rate of 59%, respectively (Narida et al, 2022). A higher striking rate would produce more successful rewarming samples.…”

“…The copyright holder for this preprint this version posted January 5, 2023. ; https://doi.org/10.1101/2023.01.04.521297 doi: bioRxiv preprint striking accuracy of more than 90%, furthermore, the sample is successfully rewarmed with a highest rate of 62% (Table 1). In this work, we built upon our previous work (Narida et al, 2022) and describe the cryo-jig A and B were operated together to be aligned to hit the same spot, however, combining this device required discretional jig tool (cryo-jig A) and jig holder (cryo-jig B) position and assembling the cryo-stick to reach higher laser striking accuracy and successful rewarming. To address this issue, we focused on developing alignment and bonding techniques for special cryo-jig which were assembling jig tool and jig holder in one piece.…”

“…Other notable achievements include the first successful cryopreservation of coral larvae with the presence of Symbiodiniaceae in larvae from Seriatopora caliendrum and Pocillopora verrucosa that have vertical transmission of algal symbionts were able to settle after vitrification and laser warming (Cirino et al, 2019(Cirino et al, , 2021. Furthermore, the long term cryo-storge after vitrification and for the following laser warming can only be achieved using our customized device which was first attempted to be developed in our prior study (Narida et al, 2022), however, this device required two jigs fitting positions and assembly Cryotop procedures to conduct high laser striking accuracy and effective rewarming. It is an urgent need to improve the efficiency and sustainability of innovative solutions for the future long-term cryobanking.…”

“…Thus according recent work from our laboratory has shown that two types of cryo-jig (cryo-jig A and B) were utilized to load and warm the sample for the vitrification and laser warming method, and the cryo-jig had a laser striking accuracy of 76% (Narida et al, 2022). It might be caused by imperfect alignment of laser machine with the jig axis circle, which was not centered on the origin.…”

First successful production of adult corals derived from cryopreserved larvae

“…These characteristics make coral planula larvae excellent candidates for cryobanking. With the advent of cryopreservation techniques, novel devices for vitrification that enable long-term storage and cryobanking can be used to laserwarm coral samples; thus, planulae are a suitable coral biomaterial for cryopreservation (Narida et al, 2022a;Lin et al, 2023). The aim of this study was to vitrify and laser-warm S. pistillata planulae to allow for feasible settlement, post-settlement survival and the production of adult corals.…”