Cryopreservation is a valuable tool for the long-term conservation of threatened and valuable plant species. Cryo-storage at Kings Park has been used since the early 1990s to provide a long-term conservation collection for species that are threatened in their native habitat, with cryopreservation used to store a wide range of plant tissue types including seeds, tissue cultured shoots and callus as well as symbiotic mycorhyzial fungi for orchid seeds. However, the process of cryopreserving a species imposes various stresses, including ice formation, oxidative stress, mitochondrial damage, and solute toxicity, all of which can limit survival rates after cryopreservation. Cryobiotechnology aims to understand and mitigate these cryo-stresses, resulting in the continued development of new and improved cryopreservation protocols. Research conducted at Kings Park Science using in vitro cultures and shoot tip material has cryopreserved 37 of our rare and threatened species, with a range of cryobiology tools used to help develop the successful cryopreservation protocols. This has included work on optimising the cryogenic protocols, thermal analysis to understand ice formation and effectiveness of the cryoprotective agents used, lipidomics studies to assess membrane compositions and membrane leakage, changes to the redox environment and the effect cryopreservation has on mitochondrial function.