A Fully Automated Robotic System for Microinjection of Zebrafish Embryos

Wang, Wenhui; Liu, Xinyu; Gelinas, Danielle; Ciruna, Brian; Sun, Yu

doi:10.1371/journal.pone.0000862

Cited by 241 publications

(126 citation statements)

References 21 publications

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“…Robotics are being developed to allow medium-and high-throughput screening (e.g. Wang et al, 2007). Finally, many human disease-related genes have orthologues in the zebrafish 2 .…”

mentioning

confidence: 99%

Zebrafish development and regeneration: new tools for biomedical research

Brittijn

Duivesteijn²,

Belmamoune³

et al. 2009

Int. J. Dev. Biol.

148

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Basic research in pattern formation is concerned with the generation of phenotypes and tissues. It can therefore lead to new tools for medical research. These include phenotypic screening assays, applications in tissue engineering, as well as general advances in biomedical knowledge. Our aim here is to discuss this emerging field with special reference to tools based on zebrafish developmental biology. We describe phenotypic screening assays being developed in our own and other labs. Our assays involve: (i) systemic or local administration of a test compound or drug to zebrafish in vivo; (ii) the subsequent detection or "readout" of a defined phenotypic change. A positive readout may result from binding of the test compound to a molecular target involved in a developmental pathway. We present preliminary data on assays for compounds that modulate skeletal patterning, bone turnover, immune responses, inflammation and early-life stress. The assays use live zebrafish embryos and larvae as well as adult fish undergoing caudal fin regeneration. We describe proof-of-concept studies on the localised targeting of compounds into regeneration blastemas using microcarriers. Zebrafish are cheaper to maintain than rodents, produce large numbers of transparent eggs, and some zebrafish assays could be scaled-up into medium and high throughput screens. However, advances in automation and imaging are required. Zebrafish cannot replace mammalian models in the drug development pipeline. Nevertheless, they can provide a cost-effective bridge between cell-based assays and mammalian whole-organism models. KEY WORDS: Danio rerio, zebrafish, high-throughput screening, high-content screeningThis article is part of a special journal issue on "Pattern Formation". In this context, it seems appropriate to ask: 'what benefits to society can come from research into pattern formation?' The answer depends on how "pattern formation" is defined. For our purposes, it includes developmental mechanisms that generate Int. J. Dev. Biol. 53: 835-850 (2009) doi: 10.1387/ijdb.082615sb One is the field of human congenital malformations (the pres-836 S. A. Brittijn et al. ence at birth of anatomical abnormalities). Good examples are human malformations of fingers and toes, some cases of which are linked to mutations in the Hox family of developmental patterning genes (Akarsu et al., 1996;Goodman et al., 1997;Goodman, 2002). Teratology -the study of agents that cause congenital malformations -can be seen as applied pattern formation research. Tissue engineering, a medical research discipline that aims to create replacement tissues for patients who have suffered injury or surgical removal of tissues, is another area where knowledge of pattern formation can be applied. To date, the commonest approach to tissue engineering is to culture autologous cells on a synthetic template (Silva and Mooney, 2004). In the future, however, it may be possible to promote pattern formation by these cells so that they generate selforganised tissues or structures ...

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“…Robotics are being developed to allow medium-and high-throughput screening (e.g. Wang et al, 2007). Finally, many human disease-related genes have orthologues in the zebrafish 2 .…”

mentioning

confidence: 99%

Zebrafish development and regeneration: new tools for biomedical research

Brittijn

Duivesteijn²,

Belmamoune³

et al. 2009

Int. J. Dev. Biol.

148

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“…These data have been used to propose a theoretical model to study and characterize the mechanical properties of biological cells, and to find the relationship between the deformation of a cell and the associated force (Sun et al 2003;Tan et al 2008). There have been many research efforts and developments directed toward automation of microinjection (Li, Zong, and Bi 2001;Kumar, Kapoor, and Taylor 2003;Matsuoka et al 2005;Mattos et al 2007;Wang, Liu, and Sun 2009;Huang et al 2009;Wang et al 2007;Yu and Nelson 2001;Ammi and Ferreira 2006). However relatively few research efforts have addressed the aforementioned issue.…”

Section: Introductionmentioning

confidence: 99%

A Vision-Based Methodology to Dynamically Track and Describe Cell Deformation during Cell Micromanipulation

Karimirad

Shirinzadeh

Yan

et al. 2013

International Journal of Optomechatronics

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The main objective of this article is to mechanize the procedure of tracking and describing the various phases of deformation of a biological circular cell during micromanipulation. The devised vision-based methodology provides a real-time strategy to track and describe the cell deformation by extracting a geometric feature called dimple angle. An algorithm based on Snake was established to acquire the boundary of the indenting cell and measure the aforementioned feature. Micromanipulation experiments were conducted for zebrafish embryos. Experimental results were used to characterize the deformation of the manipulating embryo by the devised geometric parameter. The results demonstrated the high capability of the methodology. The proposed method is applicable to the micromanipulation of other circular biological embryos such as injection of the mouse oocyte/embryo.

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“…Among these techniques, machine vision with a motorized stage or micromanipulator microinjection is effective in delivering macromolecules of a with a high precision positioning resolution (e.g., 40nm) to soluble or insoluble compound into almost any cell type with construct a fully automated cell injection system. high rates of cell viability [8], involving a glass micropipette, Recently, a first-of-its-kind fully automated zebrafish which is manipulated to poke cells and deposit materials embryo injection system was developed [19]. The system is inside.…”

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confidence: 99%

Machine Vision and Image Processing for Automated Cell Injection

Wang

Hewett

Hann

et al. 2008

2008 IEEE/ASME International Conference on Mechtronic and Embedded Systems and Applications

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Abstract-This paper presents image processing algorithms systems [13][14][15] and many tele-operated systems [16][17][18], to for cell structure recognition, which provides the desired name just a few. These systems are limited in throughput and deposition destinations without human interference for an reproducibility because operator input (e.g., locating features automated cell injection system. Adherent cells (endothelial and destinations) or operator involvement (e.g., switching cells) are the main focus. The surface and shadow information . . of the nucleoli of endothelial cells is used to extract their locations, which subsequently produce a desired deposition required. A major holdup is the need for operator input and destination inside the nucleus by Delaunay triangulation. 436 oversight in selecting or identifying specific target locations nucleoli were 92% correctly recognized, paving the way for an or/and cells. automated adherent cell injection system to be developed.

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A Fully Automated Robotic System for Microinjection of Zebrafish Embryos

Cited by 241 publications

References 21 publications

Zebrafish development and regeneration: new tools for biomedical research

Zebrafish development and regeneration: new tools for biomedical research

A Vision-Based Methodology to Dynamically Track and Describe Cell Deformation during Cell Micromanipulation

Machine Vision and Image Processing for Automated Cell Injection

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