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Published October 9, 2018 | Submitted
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3D-printable tools for developmental biology: Improving embryo injection and screening techniques through 3D-printing technology

Abstract

Developmental biology requires rapid embryo injections and screening. We applied new affordable high-resolution 3D-printing to create five easily modifiable stamp-mold tools that greatly increase injection and screening speed, while simultaneously reducing the harmful aspects of these processes. We designed two stamps that use different approaches to improve the injection efficiency for two different types of embryo, first for embryos from the snail Crepidula fornicata, and second, for those from the spider Parasteatoda tepidariorum. Both drastically improved injection speeds and embryo survival rates, even in novice hands. The other three tools were designed for rapid side-by-side organism orientating and comparison. The first screening tool allows for optimal imaging in Xenopus laevis tadpoles, while the second and third facilitate rapid high-throughput screening of Xenopus tropicalis tadpoles and Danio rerio juveniles, respectively. These designs can act as templates for many injection or screening applications.

Additional Information

The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. The authors want to thank the Marine Biological Laboratory (MBL), the Woods Hole embryology community, and the embryology course directors Richard Behringer and Alejandro Sánchez-Alvarado, for supporting the project and for providing the 3D-printer, injection systems, and the Crepidula fornicata adults. Thank you FormLabs, for lending the 3D-printer Form2 to the embryology course. Thank you, Detlev Arendt and the annelids module at MBL, for inspiring the snail injection tool. Thank you, Jonathan J. Henry, for sharing 3D-printing experience with us at MBL, printing some of the versions of stamps for us, and for being our expert hands injecting Crepidula zigotes. We thank the Henry lab, especially Kimberly Perry, for providing the Xenopus laevis tadpoles and their advice about their handling. Thank you, Helen Willsey, and Christian Bonato for providing us valuable feedback after using the high-throughput tadpole imaging device and the injection system for spiders, respectively, and sharing pictures for this manuscript. Thank you to Mariam Hamichi from Martín-Belmonte lab (CBMSO-Spain) and David Sánchez from Guerrero lab (CBMSO-Spain) for advising on the Zebrafish design and providing us the Zebrafish material. Finally, the authors want to thank their advisors, Cristina Grande and Lea Goentoro, for reviewing the manuscript and their advice. This research was funded by the Ministerio de Economía y Competitividad of Spain, PhD fellow BES-2012-052214 and short-term stays program EEBB-1-16-11411 to M.T.-G; and The Embryology Program at Woods Hole through the Helmsley Charitable Trust, Lorus J. & Margery J. Milne Scholarship, and The Society for Developmental Biology to M.J.A. R.M.H is a Senior Associate Dean of Biological Sciences in Berkeley and actual professor at the Embryology course at the MBL in Woods-Hole. The authors have declared that no competing interests exist.

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Additional details

Created:
August 19, 2023
Modified:
October 18, 2023