These prototypes were developed as open-hardware vitrification devices for sperm cryopreservation that can be integrated with existing straw platforms. The open-hardware Vitrification Device for French Straws (VD-FS) is low-cost, customizable, 3-D printable, standardized, and allows long-term sample storage and identification. The feasibility was shown for vitrifying and storing samples with multiple configurations. The results can be improved by community efforts in design alternation and evaluation of various vitrification solutions. This is the first complete open-hardware vitrification device that can be integrated with existing French-straw storage systems, providing a foundation for future community-level modifications and improvements.
Details for the development and testing can be found in the publication: Nolan Tiersch, Jacqueline Paulson, Yue Liu, and Terrence R. Tiersch. "A 3-D Printed Vitrification Device Integrated with French Straws." Hardware X (2022, https://doi.org/10.1016/j.ohx.2022.e00366). This data set includes ‘stl’ files that can be used for 3D printing (please download from Mendeley database https://data.mendeley.com/datasets/b7yy72wcxx/2), and ‘ipt’ files that can be used for design modifications in Autodesk Inventor or Fusion 360.
(1) Vitrification wands: Vitrification quality is inversely related to sample volume, because higher thermal mass leads to lower cooling rates. Thus, vitrification wands were designed with multiple configurations that differed by loop length and height (different number of layers in 3-D printing slicing). The nominal height per layer was 0.2 mm when printed with 0.4-mm printer nozzles. Loop configurations included three lengths (Short:15.4 mm, intermediate:16.6 mm, and long:24.4 mm) and three heights (1 layers, 2 layers, 3 layers). The total length of the wands was identical to French straws to fit within standard cryogenic goblet storage containers. Wands were printed with polylactic acid (PLA) thermoplastic filament, widely used for 3D printing, and can withstand liquid nitrogen temperatures.
(2) Jig apparatus for creation of protective sleeves: A self-contained jig apparatus was designed to size, indent, and cut a standard 0.5-mL cryopreservation straw (IMV Technologies, L'Aigle, France) into two protective sleeves. This apparatus consisted of a dock, a stylus, and a stencil. The stylus featured a tapered tip to produce a defined indentation on French straws that would slide along a groove fabricated in the vitrification wands. The stencil ensured correct positioning of the French straws within the jig and facilitated localization of the indentation. The stylus enabled positioning of a razor blade, stylus, and stencil during sleeve making. The stylus and stencil were self-contained by insertion into the dock after use.
Development by the Aquatic Germplasm and Genetic Resources Center (AGGRC) at the Louisiana State University Agricultural Center. Original design by Nolan Tiersch, Jacqueline Paulson, Yue Liu, and Terrence Tiersch, in partnership with the International Zebrafish Resource Center (ZIRC) and USDA National Animal Germplasm Program (NAGP). This work was funded in part by the NIH Office of Research Infrastructure Program (ORIP). This device is part of a multi-year project to develop a series of open hardware to support development of standardized and reproducible tools for the research communities that use aquatic models to study human diseases.
Please visit our website (aggrc.com) to learn more about our work.
