Standardized Assessment of Thin‐film Vitrification for Aquatic Species

Tiersch, Nolan J.; Tiersch, Terrence R.

doi:10.1080/15222055.2017.1339153

Cited by 15 publications

(17 citation statements)

References 12 publications

(19 reference statements)

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“…The maximum time to assess a sample was set at 2.5 s to ensure accurate classifications before thawing. This required a trained assessor 8 with experience in handling the devices and classification of the test solutions. A walk-in refrigerated room held at 4°C-7°C with 65%-70% relative humidity was used to assess the samples.…”

Section: Classification Of Vitrification Qualitymentioning

confidence: 99%

“…A 3D printed pedestal was used for a visual method of classifying vitrification quality. 8 Loaded loops containing films were plunged vertically into liquid nitrogen and held stationary until all bubbling stopped. After the films were frozen, the entire device was stored in liquid nitrogen for at least 1 h. Frozen films were assigned a classification number: zero (opaque, or abundant crystalline ice formation); one (translucent, or partial vitrification), or two (transparent, or substantial vitrification, glass).…”

Section: Classification Of Vitrification Qualitymentioning

confidence: 99%

“…Representative images of frozen films in prototype loops (Long, PLA) to illustrate the assessment classifications. 8 Frozen films were assigned a classification number: zero (opaque, or abundant crystalline ice formation); one (translucent, or partial vitrification); or two (transparent, or substantial vitrification, glass). ''Film fractures'' were recorded when visible cracks disabled the ability to properly assign a numerical classification.…”

Section: Figmentioning

confidence: 99%

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Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species

2019

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Vitrification is a method of cryopreservation that freezes samples rapidly, while forming an amorphous solid (''glass''), typically in small (lL) volumes. The goal of this project was to create, by three-dimensional (3D) printing, open vitrification devices based on an elliptical loop that could be efficiently used and stored. Vitrification efforts can benefit from the application of 3D printing, and to begin integration of this technology, we addressed four main variables: thermoplastic filament type, loop length, loop height, and method of loading. Our objectives were to: (1) design vitrification loops with varied dimensions; (2) print prototype loops for testing; (3) evaluate loading methods for the devices; and (4) classify vitrification responses to multiple device configurations. The various configurations were designed digitally using 3D CAD (Computer Aided Design) software, and prototype devices were produced with MakerBot Ò 3D printers. The thermoplastic filaments used to produce devices were acrylonitrile butadiene styrene (ABS) and polylactic acid (PLA). Vitrification devices were characterized by the film volumes formed with different methods of loading (pipetting or submersion). Frozen films were classified to determine vitrification quality: zero (opaque, or abundant crystalline ice formation); one (translucent, or partial vitrification), or two (transparent, or substantial vitrification, glass). A published vitrification solution was used to conduct experiments. Loading by pipetting formed frozen films more reliably than by submersion, but submersion yielded fewer filling problems and was more rapid. The loop designs that yielded the highest levels of vitrification enabled rapid transfer of heat, and most often were characterized as being longer and consisting of fewer layers (height). 3D printing can assist standardization of vitrification methods and research, yet can also provide the ability to quickly design and fabricate custom devices when needed.

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Section: Classification Of Vitrification Qualitymentioning

confidence: 99%

Section: Classification Of Vitrification Qualitymentioning

confidence: 99%

Section: Figmentioning

confidence: 99%

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Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species

2019

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“…Stock centers also present opportunities for coordinating the testing and utilization of specialized devices that can be used to standardize cryopreservation. Certain uncontrolled or variable activities in the cryopreservation process, such as producing reproducible cooling rates across hundreds of separate users (e.g., Hu et al 2017), can be addressed by widespread adoption of standardized devices in combination with accessible training (e.g., Scherr et al 2015; Tiersch and Tiersch 2017). Such devices would follow a development pathway from innovation to customization to standardization.…”

Section: Recommendations For Standardization and Harmonizationmentioning

confidence: 99%

Addressing Reproducibility in Cryopreservation, and Considerations Necessary for Commercialization and Community Development in Support of Genetic Resources of Aquatic Species

Torres

Tiersch

2018

J World Aquaculture Soc

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For the past six decades a repeated cycle of developing new cryopreservation protocols or simply reinventing them to counteract a lack of reproducibility has led to hundreds of published studies that have offered little to the establishment of a genetic resources community for aquatic species. This has hampered repository development and inhibited industrial application. Most protocols were developed without standardized approaches, leading to irreproducible studies and questionable or meaningless comparisons. Thus cryopreservation of germplasm in aquatic species would greatly benefit from strategies to facilitate reproducibility. Our objectives were to: (1) identify major sources of irreproducibility across research, small-scale, repository, and commercial-scale development levels, (2) provide recommendations to address reproducibility challenges, and (3) offer suggestions on how researchers can directly influence commercial development and application of cryopreservation research. Sources of irreproducibility include lack of standardized procedural approaches, lack of standardized terminology, and lack of reporting guidelines. To address these challenges, we propose implementation of standard operating procedures (SOP), support of stock centers and internet content for development of training programs, and strengthening of the role of scientific journals and reviewers in reducing the frequency of irreproducible outcomes. Reproducibility is the foundation for quality management programs and product reliability, and therefore standardization is necessary to assure efficient transition to commercial-scale application and repository development. Progress can only be possible through community-based approaches focused on coalescence and consensus of disparate groups involved in aquatic species cryopreservation and management of genetic resources.

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“…Meanwhile, the emergence of fused deposition modeling (FDM) three-dimensional (3-D) printing has revolutionized prototyping and fabrication in manufacturing and research [6]. The technology provides an inexpensive, simple, and portable fabrication option with high reproducibility, and has been shown to have potential applications in biotechnology and engineering including cryobiology [18,19,21]. Different thermoplastic materials can be used in 3-D printing.…”

Section: Introductionmentioning

confidence: 99%

Three-dimensional printing can provide customizable probes for sensing and monitoring in cryobiology applications

2019

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Cryopreservation has been recognized as a powerful tool for long-term preservation of genetic resources. However, the outcomes of cryopreservation by different user groups often vary due to inconsistency in procedures and freezing equipment. Herein, we report on the feasibility of providing customizable sensing probes with three-dimensional (3-D) printing to monitor cryopreservation phenomena. The objectives were to: 1) introduce 3-D printing as a fabrication method for developing customizable probes to be used in cryogenic applications; 2) design and fabricate an example of a 3-D printed sensing probe and multiplexer capable of detecting phasechange phenomena based on quantitative data regarding sample electrical resistance and temperature, and 3) demonstrate the sensing platform in cryopreservation conditions and in combination with a custom-made 3-D printed freezing device. The sensing probe developed was designed to fit within standard 0.5-ml French straws. Phase-transition phenomena were detected by analyzing electrical resistance changes. The quantitative data from this device in conjugation with a 3-D printed freezer rack provided cryopreservation capability with high reproducibility and offered an alternative to expensive programmable freezers. The use of 3-D printing provided flexibility to develop new sensing probes or modify existing designs based on specific needs. After initial prototyping, fabrication, and testing of 3-D printed sensing probes, particularly useful designs can lead to reduction of variation in performing standardized cryopreservation protocols.

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Standardized Assessment of Thin‐film Vitrification for Aquatic Species

Cited by 15 publications

References 12 publications

Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species

Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species

Addressing Reproducibility in Cryopreservation, and Considerations Necessary for Commercialization and Community Development in Support of Genetic Resources of Aquatic Species

Three-dimensional printing can provide customizable probes for sensing and monitoring in cryobiology applications

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