As an important embodiment of biomanipulation, injection of foreign materials (e.g., DNA, RNAi, sperm, protein, and drug compounds) into individual cells has significant implications in genetics, transgenics, assisted reproduction, and drug discovery. This paper presents a microrobotic system for fully automated zebrafish embryo injection, which overcomes the problems inherent in manual operation, such as human fatigue and large variations in success rates due to poor reproducibility. Based on computer vision and motion control, the microrobotic system performs injection at a speed of 15 zebrafish embryos (chorion unremoved) per minute, with a survival rate of 98% (n = 350 embryos), a success rate of 99% (n = 350 embryos), and a phenotypic rate of 98.5% (n = 210 embryos). The sample immobilization technique and microrobotic control method are applicable to other biological injection applications such as the injection of mouse oocytes/embryos and Drosophila embryos to enable high-throughput biological and pharmaceutical research.
Sundaram J, Mellein BR, Mitragotri S (2004) An experimental and theoretical analysis of ultrasound-induced permeabilization of cell membranes. Biophys J 87: 1013–1033.
Chun K, Hashiguchi G, Toshiyoshi H, Fujita H, Kikuchi Y, et al. (1999) An array of hollow micro-capillaries for the controlled injection of genetic materials into animal/plant cells. Proc IEEE International Conference on Micro Electro Mechanical Systems (MEMS'1999): 406–411.
Zappe S, Fish M, Scott MP, Solgaard O (2006) Automated MEMS-based drosophila embryo injection system for high-throughput RNAi screens. Lab Chip 6: 1012–1019.
Kobayashi K, Kato K, Saga M, Yamane M, Rothman C, et al. (1992) Subzonal insemination of a single mouse spermatozoon with a personal computer-controlled micromanipulation system. Mol Reprod Dev 33: 81–88.
Kumar R, Kapoor A, Taylor RH (2003) Preliminary experiments in robot/human cooperative microinjection. Proc IEEE International Conference on Intelligent Robots and Systems (IROS'2003): 3186–3191.
Matsuoka H, Komazaki T, Mukai Y, Shibusawa M, Akane H, et al. (2005) High throughput easy microinjection with a single-cell manipulation supporting robot. J Biotechnol 116: 185–194.
Mattos L, Grant E, Thresher R, Kluckman K (2007) New developments towards automated blastocyst microinjections. Proc IEEE International Conference on Robotics and Automation (ICRA'2007):
Pillarisetti A, Pekarev M, Brooks AD, Desai JP (2006) Evaluating the role of force feedback for biomanipulation tasks. Proc Symposium on Haptic Interfaces for Virtual Environment and Teleoperator Systems (HAPTICS'2006):
Kim DH, Hwang CN, Sun Y, Lee SH, Kim B, et al. (2006) Mechanical analysis of chorion softening in prehatching stages of zebrafish embryos. IEEE T Nanobiosci 5: 89–94.
Langenau DM, Keefe MD, Storer NY, Guyon JR, Kutok JL, et al. (2007) Effects of RAS on the genesis of embryonal rhabdomyosarcoma. Gene Dev 21: 1382–1395.
Schulte-Merker S, van Eeden FJ, Halpern ME, Kimmel CB, Nusslein-Volhard C (1994) no tail (ntl) is the zebrafish homologue of the mouse T (Brachyury) gene. Development 120: 1009–1015.
