3-D printed customizable vitrification devices for preservation of genetic resources of aquatic species

Tiersch, Connor J.; Liu, Yue; Tiersch, Terrence R.; Monroe, W. Todd

doi:10.1016/j.aquaeng.2020.102097

Cited by 14 publications

(18 citation statements)

References 31 publications

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“…With the rising popularity of low-cost resin printers, advances in LCD technologies will likely address these problems in the near future, and bring in next-generation resolution capabilities (e.g., 8K resolution). Approaches such as these can be especially powerful when presented in the form of open hardware by functioning as force multipliers in addressing large, intractable, global problems, such as ensuring quality management for germplasm repository development in aquatic species [43,44].…”

Section: Discussionmentioning

confidence: 99%

Low-Cost Resin 3-D Printing for Rapid Prototyping of Microdevices: Opportunities for Supporting Aquatic Germplasm Repositories

Zuchowicz

Belgodere

Liu

et al. 2022

Fishes

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Germplasm repositories can benefit sustainable aquaculture by supporting genetic improvement, assisted reproduction, and management of valuable genetic resources. Lack of reliable quality management tools has impeded repository development in the past several decades. Microfabricated open-hardware devices have emerged as a new approach to assist repository development by providing standardized quality assessment capabilities to enable routine quality control. However, prototyping of microfabricated devices (microdevices) traditionally relies on photolithography techniques that are costly, time intensive, and accessible only through specialized engineering laboratories. Although resin 3-D printing has been introduced into the microfabrication domain, existing publications focus on customized or high-cost (>thousands of USD) printers. The goal of this report was to identify and call attention to the emerging opportunities to support innovation in microfabrication by use of low-cost (<USD 350) resin 3-D printing for rapid prototyping. We demonstrate that low-cost mask-based stereolithography (MSLA) 3-D printers with straightforward modifications can provide fabrication quality that approaches traditional photolithography techniques. For example, reliable feature sizes of 20 µm with dimensional discrepancy of <4% for lateral dimensions and <5% for vertical dimensions were fabricated with a consumer-level MSLA printers. In addition, alterations made to pre-processing, post-processing, and printer configuration steps improved print quality as demonstrated in objects with sharper edges and smoother surfaces. The prototyping time and cost of resin 3-D printing (3 h with USD 0.5/prototype) were considerably lower than those of traditional photolithography (5 d with USD 80/prototype). With the rapid advance of consumer-grade printers, resin 3-D printing can revolutionize rapid prototyping approaches for microdevices in the near future, facilitating participation in interdisciplinary development of innovative hardware to support germplasm repository development for aquatic species.

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Section: Discussionmentioning

confidence: 99%

Low-Cost Resin 3-D Printing for Rapid Prototyping of Microdevices: Opportunities for Supporting Aquatic Germplasm Repositories

Zuchowicz

Belgodere

Liu

et al. 2022

Fishes

Self Cite

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“…Specialized equipment was required to produce the master wafer molds. However, the evaluated designs can be used in open-hardware applications [45][46][47]. Advancements in 3-D printing have allowed for fabrication of microfluidic devices with consumer-level machines that cost <USD 400 [22,23].…”

Section: Discussionmentioning

confidence: 99%

Development of a Single-Piece Sperm Counting Chamber (SSCC) for Aquatic Species

Belgodere

Liu

Reich

et al. 2022

Fishes

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Accurate determination of sperm concentration in aquatic species is important for assisted reproduction and cryopreservation, yet is challenging as current counting methods are costly or not suitable for many species. The goal of this work was to develop a simple (single-piece and single-layer photolithography) sperm counting chamber (SSCC) for aquatic species. Goldfish (Carassius auratus) and zebrafish (Danio rerio) sperm were used for evaluation in the device, which was created with soft lithography. Four designs with different geometries were evaluated for counting accuracy. Open-corner and open-midpoint designs were the most accurate with no significant differences (P > 0.05) for most of the target sperm concentrations (0.5–1.0 × 108 cells/mL). The open-corner design was not significantly different from the Makler® counting chamber intended for human sperm cells (P = 0.6) but was significantly different from a hemocytometer (P < 0.001) intended for other cell sizes. Material cost of device production was USD 16 per unit, including photolithography supplies, glass slide and coverslip, and polydimethylsiloxane. The cost can be reduced to USD 2 per unit with repeated wafer casts. This device could be further refined for resin 3-D printing and sharing via open-hardware approaches and modified to best suit species specific applications.

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“…In addition, design files can be shared on a number of sites (e.g., Thingiverse, NIH 3D Print Exchange, and Github) for sharing and distribution. In this way, devices used in cryopreservation and repository development can be developed, shared, and standardized within research communities, greatly reducing costs of cryopreservation, and making reliable methods widely available [ 36 ]. However, distributed production systems such as this must be accompanied by quality control (QC) and quality assurance (QA) programs to ensure that samples meet minimum thresholds for repository use [ 37 , 38 ].…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of Hydractinia symbiolongicarpus Sperm to Support Community-Based Repository Development for Preservation of Genetic Resources

Huene

Koch

Arregui

et al. 2022

Animals

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Hydractinia symbiolongicarpus is an emerging model organism in which cutting-edge genomic tools and resources are being developed for use in a growing number of research fields. One limitation of this model system is the lack of long-term storage for genetic resources. The goal of this study was to establish a generalizable cryopreservation approach for Hydractinia that would support future repository development for other cnidarian species. Specific objectives were to: (1) characterize basic parameters related to sperm quality; (2) develop a generalizable approach for sperm collection; (3) assess the feasibility of in vitro fertilization (IVF) with sperm after refrigerated storage; (4) assess the feasibility of IVF with sperm cryopreserved with various sperm concentrations; (5) evaluate feasibility of cryopreservation with various freezing conditions, and (6) explore the feasibility of cryopreservation by use of a 3-D printed open-hardware (CryoKit) device. Animal husbandry and sperm collection were facilitated by use of 3-D printed open hardware. Hydractinia sperm at a concentration of 2 × 107 cells/mL stored at 4 °C for 6 d were able to achieve 50% fertilization rate. It appeared that relatively higher sperm concentration (>5 × 107 cells/mL) for cryopreservation could promote fertilization. A fertilization rate of 41–69% was observed using sperm equilibrated with 5, 10, or 15% (v/v) cryoprotectant (dimethyl sulfoxide or methanol) for 20 min, cooled at a rate of 5, 10, or 20 °C/min from 4 °C to −80 °C, at a cell concentration of 108/mL, in 0.25 mL French straws. Samples cryopreserved with the CryoKit produced a fertilization rate of 72–82%. Establishing repository capabilities for the Hydractinia research community will be essential for future development, maintenance, protection, and distribution of genetic resources. More broadly, these generalizable approaches can be used as a model to develop germplasm repositories for other cnidarian species.

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3-D printed customizable vitrification devices for preservation of genetic resources of aquatic species

Cited by 14 publications

References 31 publications

Low-Cost Resin 3-D Printing for Rapid Prototyping of Microdevices: Opportunities for Supporting Aquatic Germplasm Repositories

Low-Cost Resin 3-D Printing for Rapid Prototyping of Microdevices: Opportunities for Supporting Aquatic Germplasm Repositories

Development of a Single-Piece Sperm Counting Chamber (SSCC) for Aquatic Species

Cryopreservation of Hydractinia symbiolongicarpus Sperm to Support Community-Based Repository Development for Preservation of Genetic Resources

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