Device:

AGGRC Strand Centi-Sizer v4.0.1 (Parametric Uniform 1-cm Cut) part of the Strand Cryopreservation Shuttle System (SCSS)

Description: 

Prior to open-hardware device development, Aplysia egg masses were dissected into strands by use of a scalpel or scissors, measured to approximate size by eye or tape guide, and cut again by use of a scalpel or razor blade. The approach mirrored the traditional use of improvised, self-limiting, solutions to research problems. This process was time consuming and required excessive person-hours (# of workers × amount of time per person to accomplish a task) depending on the complexity of an egg mass (e.g., how tightly it was wound) and how many strand pieces were required. The Strand Centi-Sizer was developed to increase efficiency and accuracy of cutting Aplysia egg strands in precisely sized, 1-cm pieces. The Strand Centi-Sizer was printed by use of a fused filament fabrication (FFF) 3-D printer with polylactic acid (PLA) thermoplastic filament (ZYLtech, Houston, Texas). The device was 50 mm × 120 mm × 7 mm and had two parallel grooves (spaced 21.4 mm on center) and multiple cutting channels, perpendicular to the grooves, spaced according to the length of egg strand required by the user. This version is appropriate for cutting 1-cm strands.

Fusion360 and other files can be downloaded here: https://zenodo.org/record/8033755.

Development by the Aquatic Germplasm and Genetic Resources Center (AGGRC) at the Louisiana State University Agricultural Center. Original design by Allyssa Oune. Designed in partnership with the National Resource for Aplysia. This work was funded in part by NIH Office of Research Infrastructure Program (ORIP). 

Device:

AGGRC Strand Centi-Sizer v4.0.1 (Parametric Mixed Cut) part of the Strand Cryopreservation Shuttle System (SCSS)

Description:

Prior to open-hardware device development, Aplysia egg masses were dissected into strands by use of a scalpel or scissors, measured to approximate size by eye or tape guide, and cut again by use of a scalpel or razor blade. The approach mirrored the traditional use of improvised, self-limiting, solutions to research problems. This process was time consuming and required excessive person-hours (# of workers × amount of time per person to accomplish a task) depending on the complexity of an egg mass (e.g., how tightly it was wound) and how many strand pieces were required. The Strand Centi-Sizer was developed to increase efficiency and accuracy of cutting Aplysia egg strands in precisely sized, 1-cm pieces. The Strand Centi-Sizer was printed by use of a fused filament fabrication (FFF) 3-D printer with polylactic acid (PLA) thermoplastic filament (ZYLtech, Houston, Texas). The device was 50 mm × 120 mm × 7 mm and had two parallel grooves (spaced 21.4 mm on center) and multiple cutting channels, perpendicular to the grooves, spaced according to the length of egg strand required by the user. This version is appropriate for cutting 1-cm or 0.25-cm strands.

Fusion360 and other files can be downloaded here: https://zenodo.org/record/8033755.

Development by the Aquatic Germplasm and Genetic Resources Center (AGGRC) at the Louisiana State University Agricultural Center. Original design by Allyssa Oune. Designed in partnership with the National Resource for Aplysia. This work was funded in part by NIH Office of Research Infrastructure Program (ORIP). 

Device:

AGGRC Strand Shuttle Cassette v6.1.16 and Aligner Sealer v2.1.4 part of the Strand Cryopreservation Shuttle System (SCSS)

Description:

Strand Shuttle Cassette

The Strand Shuttle Cassette was designed to capture and support 10 egg strand pieces as long as 1 cm for application to equilibrium cooling. Each cassette had a rectangular base (15 mm × 53 mm × 2 mm) with 10 perpendicular shuttles (Figure 4A). Each shuttle was 28.5 mm × 2 mm allowing it to fit unimpeded within a 0.5-mL French straw. Shuttles were spaced 5.40 mm on center to match the straw spacing on most 0.5 mL-compatible cryopreservation equipment. Each shuttle was connected to the base by a tapered cylinder to facilitate removal of the shuttle from the cassette. Distal to the tapered cylinder was a cage-like slot designed to hold a single egg strand piece. The slot had an open back to reduce the thermal mass surrounding the egg strand and open ends to facilitate cryoprotectant solution equilibration and shuttle movement down the length of a French straw. The most distal end of each cassette was topped with a 3-mm 15˚ conical point to assist in guiding the shuttle into a straw. The number, size, and spacing of the shuttles were customizable through edits in CAD software (Fusion360, Autodesk, San Francisco, California). In this configuration a single 0.5-mL French straw could hold three shuttles.

Aligner Sealer

The Aligner Sealer was designed to have two roles: align strands on the Strand Base and seal straws without needing heat, water (to moisten PVA sealing powder), or electrical power. The Aligner Sealer was a rectangular device (15 mm × 53 mm × 2 mm) with 10, 5-mm long cylindrical pegs (Figure 4B). Each peg had a 2.58-mm diameter ball attached 1.4 mm from the base, and each was topped with a 2-mm 15˚ conical point to assist in guiding the peg into a straw. Peg spacing aligned with the grooves on the Strand Base and the straws in the Straw Holder. The size of the sealing ball and spacing of the pegs were customizable through edits in CAD software (Fusion360, Autodesk, San Francisco, California).

Fusion360 and other files can be downloaded here: https://zenodo.org/record/8033755.

Development by the Aquatic Germplasm and Genetic Resources Center (AGGRC) at the Louisiana State University Agricultural Center. Original design by Jack Koch. Modifications by Cameron Bonds. Designed in partnership with the National Resource for Aplysia. This work was funded in part by NIH Office of Research Infrastructure Program (ORIP). 

Device:

AGGRC Strand Base v4.0.0 part of the Strand Cryopreservation Shuttle System (SCSS)

Description:

The Strand Base was designed to support egg strand pieces in preparation for loading into Strand Shuttle Cassette shuttles. The base was a rectangular device (29 mm × 65 mm × 7 mm) with 10 parallel grooves ~3.2 mm wide and ~1.8 mm deep to accommodate egg strands and shuttles (Figure 4C). Groove spacing aligned with shuttles on the Strand Shuttle Cassette and pegs on the Aligner Sealer. The depth, width, and spacing of the grooves were customizable through edits in CAD software (Fusion360, Autodesk, San Francisco, California).

Fusion360 and other files can be downloaded here: https://zenodo.org/record/8033755.

Development by the Aquatic Germplasm and Genetic Resources Center (AGGRC) at the Louisiana State University Agricultural Center. Original design by Jack Koch. Modifications by Cameron Bonds. Designed in partnership with the National Resource for Aplysia. This work was funded in part by NIH Office of Research Infrastructure Program (ORIP). 

Device:

AGGRC Straw Holder v1.8.6 part of the Strand Cryopreservation Shuttle System (SCSS)

Description:

The Straw Holder was designed to firmly hold 10, 0.5-mL French straws. The straw-holding side was a rectangle (54 mm × 65 mm × 8 mm) with 10 parallel grooves ~3.4 mm wide and ~1.9 mm deep (Figure 4D). The pressure side was a rectangle (54 mm × 65 mm × 8 mm) with no grooves. The two side were connected by a hinge. There was a locking mechanism that enabled straws to be held in place to facilitate loading of shuttles and straw sealing. The depth, width, and spacing of the grooves were customizable through edits in CAD software (Fusion360, Autodesk, San Francisco, California).

Fusion360 and other files can be downloaded here: https://zenodo.org/record/8033755.

Development by the Aquatic Germplasm and Genetic Resources Center (AGGRC) at the Louisiana State University Agricultural Center. Original design by Jack Koch. Modifications by Cameron Bonds. Designed in partnership with the National Resource for Aplysia. This work was funded in part by NIH Office of Research Infrastructure Program (ORIP).