An automated modular open-technology device to measure and adjust concentration of aquatic sperm samples for cryopreservation

Abstract

Repositories for aquatic germplasm are essential for safeguarding valuable genetic diversity for species relevant to aquaculture, biomedical research, and conservation. Development of aquatic germplasm repositories is impeded by a lack of standardization within laboratories and across the research community. Protocols for cryopreservation are often developed ad hoc and without close attention to variables, such as cell concentration, that strongly affect the success and reproducibility of cryopreservation. The wide dissemination and use of specialized tools and devices as open hardware can improve processing reliability and save costs. The goal of the present work was to develop and prototype a modular and open-technology approach to help to standardize the cell concentration of germplasm samples prior to cryopreservation. The specific objectives were to: 1) design and fabricate prototypes of the automated concentration measurement and adjustment system (CMAS), incorporating custom peristaltic pumps and optical evaluation modules, and 2) evaluate the performance of the CMAS with biological samples. Linear regression models were obtained for estimation of aquatic sperm concentration >108 cells/mL and for algae concentration > (3 × 105) cells/mL. Algae were diluted with extender medium by an automated process, resulting in a dilution precision of ±12.6% and ±6.7% in two trials, attaining means of 89% and 71% of the target cell concentration. The development of the CMAS as open technology can provide opportunities for community-level standardization in cryopreservation of aquatic germplasm and can invite new users, makers, and developers into the open-technology community. This will increase the reach and capabilities of much-needed aquatic germplasm repositories.

Keywords

• Open technology
• Concentration
• Dilution
• 3-D Printing
• Aquatic
• Cryopreservation