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海胆受精卵的高通量显微注射。

High throughput microinjections of sea urchin zygotes.

作者信息

Stepicheva Nadezda A, Song Jia L

机构信息

Department of Biological Sciences, University of Delaware.

出版信息

J Vis Exp. 2014 Jan 21(83):e50841. doi: 10.3791/50841.

Abstract

Microinjection into cells and embryos is a common technique that is used to study a wide range of biological processes. In this method a small amount of treatment solution is loaded into a microinjection needle that is used to physically inject individual immobilized cells or embryos. Despite the need for initial training to perform this procedure for high-throughput delivery, microinjection offers maximum efficiency and reproducible delivery of a wide variety of treatment solutions (including complex mixtures of samples) into cells, eggs or embryos. Applications to microinjections include delivery of DNA constructs, mRNAs, recombinant proteins, gain of function, and loss of function reagents. Fluorescent or colorimetric dye is added to the injected solution to enable instant visualization of efficient delivery as well as a tool for reliable normalization of the amount of the delivered solution. The described method enables microinjection of 100-400 sea urchin zygotes within 10-15 min.

摘要

向细胞和胚胎中进行显微注射是一种常用技术,用于研究广泛的生物学过程。在这种方法中,将少量处理溶液装入显微注射针中,该针用于物理注射单个固定的细胞或胚胎。尽管进行高通量注射操作需要初步培训,但显微注射能以最高效率将各种处理溶液(包括复杂的样品混合物)可重复地注射到细胞、卵子或胚胎中。显微注射的应用包括递送DNA构建体、mRNA、重组蛋白、功能获得和功能丧失试剂。向注射溶液中添加荧光或比色染料,以便即时观察有效递送情况,并作为可靠标准化递送溶液量的工具。所述方法能够在10至15分钟内对100 - 400个海胆受精卵进行显微注射。

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