Microinjection in C. elegans by direct penetration of elastomeric membranes.
Microinjection in C. elegans by direct penetration of elastomeric membranes.
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线虫线虫被广泛应用于基础研究和翻译研究。在许多线虫研究项目中，通过将DNA结构注入线虫的性腺来创造转基因菌株是必不可少的一步。本文介绍了一种用于进行微注射的最低限度微流控芯片的制造和使用。蠕虫被固定在一个紧密配合的微通道中，该微通道的一侧是一层薄薄的弹性膜，注射管穿过该膜到达蠕虫。吸管既不会破裂，也不会通过膜堵塞，当吸管被取出时，膜会重新密封。存活率和转化率与传统方法相似。新手用户发现，使用该设备进行注射比传统方法更容易学习。弹性体膜的直接穿透原理适用于在包括细胞、胚胎和其他小动物模型在内的各种生物体中进行微量注射。因此，它可能导致用于基础、平移和工业应用的新一代微注射系统。
The nematode worm C. elegans is widely used in basic and translational research. The creation of transgenic strains by injecting DNA constructs into the worm's gonad is an essential step in many C. elegans research projects. This paper describes the fabrication and use of a minimalist microfluidic chip for performing microinjections. The worm is immobilized in a tight-fitting microchannel, one sidewall of which is a thin elastomeric membrane through which the injection pipet penetrates to reach the worm. The pipet is neither broken nor clogged by passing through the membrane, and the membrane reseals when the pipet is withdrawn. Rates of survival and transgenesis are similar to those in the conventional method. Novice users found injections using the device easier to learn than the conventional method. The principle of direct penetration of elastomeric membranes is adaptable to microinjections in a wide range of organisms including cells, embryos, and other small animal models. It could, therefore, lead to a new generation of microinjection systems for basic, translational, and industrial applications.
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