A practical device for pinpoint delivery of molecules into multiple neurons in culture
A practical device for pinpoint delivery of molecules into multiple neurons in culture
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DOI：
10.1007/s11068-008-9021-z
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发表时间：
2006-12-01
期刊：
影响因子：
--
通讯作者：
Miyawaki, Atsushi
中科院分区：
文献类型：
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作者：
Hara, Chikako;Tateyama, Kiyohiko;Miyawaki, Atsushi
我们开发了一种将化学物质、蛋白质和核酸精确输送到培养细胞中的装置。该技术的原理是分子通过针刺造成的质膜破裂从培养基流到细胞中。 DNA 转染是通过将针尖刺入细胞核来实现的。 CellBee 装置可以连接到任何倒置显微镜上，分子输送可以与传统的活细胞成像相结合。由于针相对于目标培养细胞的位置是由计算机控制的，因此可以在 10 分钟内完成将罗丹明等分子有效输送到多达 100 个 HeLa 细胞中。此外，通过简单改变含有不同质粒的培养基，可以用多种DNA构建体转染单个培养皿内的特定靶细胞。此外，纳米级针尖能够温和地输送分子，最大限度地减少细胞损伤。该方法允许将 DNA 转染到特定的海马神经元中，而不会干扰培养物中建立的神经元回路。
We have developed a device for pinpoint delivery of chemicals, proteins, and nucleic acids into cultured cells. The principle underlying the technique is the flow of molecules from the culture medium into cells through a rupture in the plasma membrane made by a needle puncture. DNA transfection is achieved by stabbing the needle tip into the nucleus. The CellBee device can be attached to any inverted microscope, and molecular delivery can be coupled with conventional live cell imaging. Because the position of the needle relative to the targeted cultured cells is computer-controlled, efficient delivery of molecules such as rhodamine into as many as 100 HeLa cells can be completed in 10 min. Moreover, specific target cells within a single dish can be transfected with multiple DNA constructs by simple changes of culture medium containing different plasmids. In addition, the nanosized needle tip enables gentle molecular delivery, minimizing cell damage. This method permits DNA transfection into specific hippocampal neurons without disturbing neuronal circuitry established in culture.