Aquaculture America 2024

February 18 - 21, 2024

San Antonio, Texas

AN OPEN-HARDWARE APPROACH FOR REAL-TIME ELECTRICAL SENSING OF VITRIFICATION TO SUPPORT GERMPLASM REPOSITORY DEVELOPMENT FOR AQUATIC SPECIES

Pedram Hosseinzadehnamadi* , Caleb L. Gryder, Yue Liu, Terrence R. Tiersch

                                 Aquatic Germplasm and Genetic Resources Center

                                 Louisiana State University Agricultural Center

                                 Baton Rouge, LA, 70820

                                 Phosse3@lsu.edu



                                

Development of germplasm repositories for aquatic species is vital for protecting valuable genetic resources. The recovery success of the cells after cryopreservation is highly dependent on freezing and warming conditions due to potential cell damage during the process. In cryopreservation, equilibrium, and non-equilibrium (vitrification) cooling are commonly used methods. Vitrification typically employs ultra-rapid cooling (e.g., >10,000 °C/min) to produce liquid-to-solid phase change into a glass-like state that minimizes ice crystal formation. We seek to enable real-time monitoring of the vitrification process through innovative electrical sensing technologies. Traditional assessment of vitrification has primarily relied on visual observation of sample transparency, which can be subjective and unreliable. Typically, crystalline ice is opaque or translucent, whereas vitrified samples remain transparent. To make this work more accessible, it is based on open fabrication techniques such as resin 3-D printing and printed circuit boards (PCB) to develop custom probes that can be shared by the internet (Figure 1A). These probes can detect electrical impedance signals to monitor phase change (e.g., liquid to solid). Preliminary results (Figure 1B) show an increase in electrical resistance of a 20% dimethyl sulfoxide solution as it undergoes vitrification. These impedance changes reflect formation of ice and validate the feasibility of monitoring vitrification by use of prototype probes fabricated by resin printing of a substrate that could be equipped with electrical circuitry. The impedance increased during cooling reflecting vitrification at the glass transition (Tg) point without a clear release of latent heat (typically seen when cooling more slowly). This approach can offer quantifiable and precise monitoring, enhancing the reproducibility of cryopreservation through vitrification. Open fabrication technologies will enable user communities to fabricate, adapt, and improve such probes, thus contributing to development of germplasm repositories at a community level.