A Review of Automated Microinjection of Zebrafish Embryos

Abstract

Cell microinjection is a technique of precise delivery of substances into cells and is widely used for studying cell transfection, signaling pathways, and organelle functions. Microinjection of the embryos of zebrafish, the third most important animal model, has become a very useful technique in bioscience. However, factors such as the small cell size, high cell deformation tendency, and transparent zebrafish embryo membrane make the microinjection process difficult. Furthermore, this process has strict, specific requirements, such as chorion softening, avoiding contacting the first polar body, and high-precision detection. Therefore, highly accurate control and detection platforms are critical for achieving the automated microinjection of zebrafish embryos. This article reviews the latest technologies and methods used in the automated microinjection of zebrafish embryos and provides a detailed description of the current developments and applications of robotic microinjection systems. The review covers key areas related to automated embryo injection, including cell searching and location, cell position and posture adjustment, microscopic visual servoing control, sensors, actuators, puncturing mechanisms, and microinjection.

Drug Administration Routes Impact the Metabolism of a Synthetic Cannabinoid in the Zebrafish Larvae Model

“…Microinjection has been used already for decades as a way of administering compounds to fish in early life stages, and this technique is still widely used in cellular microbiological research [ 18 , 19 , 20 ]. However, most laboratories use aquatic exposure in screening campaigns with ZF larvae, thus neglecting the potentially hindered uptake and absorption of lipophilic compounds which, in turn, can lead to a rather high rate of false negatives.…”

A Review of Recent Advances in Robotic Cell Microinjection

“…As shown in Table 2, Zhao et al [17] summarized the advantages and disadvantages of current popular actuators. Among them, piezoelectric ceramic actuator has the advantages of high stiffness, large blocking force, fast response speed, and being applicable to high frequency.…”

“…These issues mean that manual methods can only be used for small number of operations for single-cell injections [15]. With the development of automatic control technology and micro/nano-positioning platform, robotic cell microinjection method has been realized, which can meet the requirements of biological sciences, to complete fast and accurate microinjection tasks of a large number of cells [16], [17].…”

Modeling Neuronal Diseases in Zebrafish in the Era of CRISPR

“…Moreover, injecting Cas protein instead of Cas mRNA could improve mutagenesis efficiency [ 61 ]. Third, these sgRNAs together with the Cas mRNA or protein are injected into fertilized embryos at the one-cell stage [ 62 , 63 ]. Injection dose should be standardized to improve efficiency and avoid phenotypes due to the toxic effect of the injection components.…”

“…Injection dose should be standardized to improve efficiency and avoid phenotypes due to the toxic effect of the injection components. Microinjection efficiency can also improve using an automated microinjection system [ 62 ]. Each injected embryo grows as a mosaic founder fish (crispant) with various InDels in the somatic cells as the nucleases cleave the target sites over the course of multiple rounds of cell division in the developing embryos [ 44 ].…”