Semester of Graduation

Fall 2021

Degree

Master of Science (MS)

Department

School of Renewable Natural Resources

Document Type

Thesis

Abstract

Repositories for aquatic germplasm can safeguard the genetic diversity of species of interest to aquaculture, research, and conservation. The development of such repositories is impeded by a lack of standardization both within laboratories and across the research community. Protocols for cryopreservation are often developed ad hoc and without close attention to variables, such as sperm concentration, that strongly affect the success and consistency of cryopreservation. The wide dissemination and use of specialized tools and devices can improve processing reliability, provide data logging, produce custom hardware to address unique problems, and save costs, time, and labor. The goal of the present work was to develop a low-cost and open-technology approach to standardize the concentration of sperm samples prior to cryopreservation. The specific objectives were to: 1) fabricate and test a peristaltic pump and optical evaluation module (POEM); 2) fabricate and test a prototype of the modular, open-technology concentration measurement and adjustment system (CMAS), which incorporated the POEM; 3) identify opportunities to extend the CMAS to microliter volumes through low-cost resin 3-D printing, and 4) identify strategies from this work that can be applied to future open fabrication efforts. The POEM and CMAS were prototyped and tested with biological samples. A relationship between optical signal and cell concentration of channel catfish (Ictalurus punctatus) sperm samples was established by linear regression. In a blind trial, cell concentrations were estimated with the POEM and correlated closely to their known concentrations (linear regression R2 = 0.9945) in a range of 1 × 108 to 4 × 109 cells/mL. The CMAS was able to estimate and adjust the concentration of a sample of the marine microalgae Tetraselmis chuii as a preparatory step for cryopreservation. To explore the possible use of the CMAS with microliter sample volumes in future work, evaluation of low-cost resin 3-D printing showed that this technology can approach conventional microfabrication techniques in feature quality and resolution. The development of the CMAS as open technology can provide opportunities for community-level standardization in cryopreservation of aquatic germplasm, invite new users, makers, and developers into the open-technology community, and increase the reach and capabilities of aquatic germplasm repositories.

Committee Chair

Tiersch, Terrence R.

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