Journal: bioRxiv

Article Title: The first proof of concept demonstration of nanowarming in coral tissue

doi: 10.1101/2023.03.16.533048

Figure Lengend Snippet: Production, cryopreservation, and recovery of single-polyp microfragments (SPMFs) from the coral Porites compressa . Corals were cut into microfragments and grown on plastic sheets, then individual polyps were excised from thin primary growth areas using a biopsy punch and allowed to heal in an incubator to form single-polyp microfragments (SPMFs). Single-polyp microfragments were pre-incubated in 0.07 mg/mL (4.5 × 10 np/m -3 ) 1064 nm resonant PEG-coated gold nanorods in FSW for 1 hour, then equilibrated with cryoprotectant solution in three steps over 4 minutes on the cryotop. The cryotop with SPMF was transferred to the cryo device for vitrification and laser nanowarming using an iWeld 980 Series, 60-joule, Nd:YAG unpolarised infrared laser (240 V, 9.0 ms, ∼ 8.2 J). Single-polyp microfragments were then rehydrated in five steps over 4 minutes, followed by recovery in a 24-well plate overnight in an incubator without light.

Article Snippet: The wooden holder with the cryo blade and SPMF was then transferred into the chamber of a bench-top iWeld 980 Series, 60 joule, Nd:YAG unpolarised infrared laser (model number 585-986-080; LaserStar Technologies Corporation, Orlando, FL, USA) and inserted into the arm of a custom-made cryo laser apparatus (Design Solutions, Inc., Chanhassen, MN, USA) for vitrification and laser nanowarming as described in Daly et al. . After 1 min exposure to the full-strength vitrification solution, the cryo apparatus was used to lower the sample into a liquid nitrogen bath for vitrification.

Techniques: Incubation