爪蟾卵母细胞:在96孔微滴板上自动注射cDNA/mRNA并随后进行TEVC记录。

K. Schnizler, Mike Küster, C. Methfessel, M. Fejtl
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引用次数: 60

摘要

膜结合的神经递质受体和离子通道是数量最多和最重要的药物靶点之一,电生理方法是研究其功能特性和对药物反应的金标准。然而,电生理测量通常由高技能的个体一次执行一个,并且由于缺乏吞吐量,二次功能筛选经常受到阻碍。因此,使用自动化程序来提高电生理技术的效率是非常有趣的。在已经描述的许多不同的电生理技术中,爪蟾卵母细胞的两个电极电压钳记录(TEVC)似乎特别适合于自动化测量系统的实施。在这里,我们描述了一个专门为此目的开发的工作站。Roboocyte是第一个(也是目前唯一可用的)仪器,可以自动执行cDNA(或mRNA)注射和随后的TEVC记录,将爪蟾卵细胞涂在标准的96孔微滴板上。本文介绍了用于药物筛选的卵母细胞表达系统的科学背景和卵母细胞的发展。然后,介绍了机器人细胞系统的一些技术细节,最后,讨论了与人工进行的实验相比,机器人细胞所获得的关于提高通量的结果。欲了解更多信息,请访问www.roboocyte.com。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The roboocyte: automated cDNA/mRNA injection and subsequent TEVC recording on Xenopus oocytes in 96-well microtiter plates.
Membrane-bound neurotransmitter receptors and ion channels are among the most numerous and important drug targets, and electrophysiological methods are the gold standard for the study of their functional properties and their response to drugs. However, electrophysiological measurements are usually performed one at a time by highly skilled individuals, and secondary functional screening is often hampered by this lack of throughput. Accordingly, the use of automated procedures to increase the efficiency of electrophysiological techniques is of great interest. Among the many different electrophysiological techniques that have been described, two electrode voltage clamp recording (TEVC) from Xenopus oocytes seems particularly suitable for the implementation of automated measurement systems. Here, we describe a workstation that was expressly developed for this purpose. The Roboocyte is the first (and the only currently available) instrument that automatically performs both cDNA (or mRNA) injection and subsequent TEVC recording on Xenopus oocytes plated in a standard 96-well microtiter plate. This paper describes the scientific background of the oocyte expression system for drug screening and the development of the Roboocyte. Then, some technical details of the Roboocyte system are presented and, finally, results obtained with the Roboocyte are discussed with regard to increased throughput compared with manually performed experiments. Further information can be obtained at www.roboocyte.com.
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