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

Vitrification and laser warming have gained popularity over the traditional convective warming techniques in cryopreservation. Laser warming is rapid with uniform effects, thus preventing ice crystal formation in samples. Contemporary laser warming studies have focused on proof-of-concept experiments. Yet, no protocols or techniques have been developed to address the problem of warming samples from long-term storage. Herein, a new approach to laser warming samples without exposing the samples to ambient temperature is introduced. The new device presented has a mean laser-hitting accuracy of 76% ± 16% and a rewarming rate of 59% ± 25% on samples with <1 μL in volume. Although these rates depend on the choice of vitrification solution and mastery of the technique, the approach described represents a successful first step toward laser warming samples from long-term cryo-storage.