Automated tracking and laser micromanipulation of motile cells -: art. no. 035105
Automated tracking and laser micromanipulation of motile cells -: art. no. 035105
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DOI：
10.1063/1.1863812
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发表时间：
2005-03-01
影响因子：
1.6
通讯作者：
Käs, J
中科院分区：
文献类型：
--
作者：
Stuhrmann, B;Gögler, M;Käs, J
对神经元生长的控制是建立明确的体外神经元网络的先决条件，作为阐明神经元间通讯的分析方法。神经元的生长是通过将近红外激光聚焦在神经细胞的前沿来引导的[A.Ehrlicher，T.Betz，B.Stuhrmann，D.Koch，V.Milner，M.G.Raizen，and J.Kas，Proc。娜塔莉。阿卡德。SCI。美国，99,16024(2002年)]。Ehrlicher报告的设置仅限于局部激光照射，并且依赖于大量的主观交互，因为激光光束只能手动操控。为了克服已报道的设置的缺点，我们开发并在此提供了一个全自动低对比度边缘检测软件包，该软件包通过快速启动激光转向装置(如声光偏转器或移动反射镜)来响应检测到的细胞形态变化，从而使实验能够在最小人为干扰的情况下进行。由此产生的辐射模式可以是空间、时间和细胞形态的任意函数，并且通过实验特定的反馈例程来计算。数据处理在1 S的数量级上重复，允许对形态变化做出快速反应。我们的程序的优点是将实时低对比度形状检测与复杂的反馈机制相结合，以及由于模块化编程概念而易于适应。在这篇文章中，我们演示了自动光学制导；然而，该软件很容易适用于其他需要设备对低对比度物体的形态变化做出自动快速反应的问题。(C)2005年美国物理研究所。
Control over neuronal growth is a prerequisite for the creation of defined in vitro neuronal networks as assays for the elucidation of interneuronal communication. Neuronal growth has been directed by focusing a near-infrared laser beam at a nerve cell's leading edge [A. Ehrlicher, T. Betz, B. Stuhrmann, D. Koch, V. Milner, M. G. Raizen, and J. Kas, Proc. Natl. Acad. Sci. U.S.A. 99, 16024 (2002)]. The setup reported by Ehrlicher was limited to local laser irradiation and relied on a great deal of subjective interaction since the laser beam could only be steered manually. To overcome the drawbacks of the reported setup, we developed and here present a fully automated low-contrast edge detection software package, which responds to detected cell morphological changes by rapidly actuating laser steering devices, such as acousto-optical deflectors or moving mirrors, thus enabling experiments with minimum human interference. The resulting radiation patterns can be arbitrary functions of space, time, and cell morphology, and are calculated by experiment specific feedback routines. Data processing is repeated on the order of 1 s allowing rapid reactions to morphological changes. The strengths of our program are the combination of real-time low contrast shape detection with complex feedback mechanisms, as well as easy adaptability due to a modular programming concept. In this article we demonstrate automated optical guidance; however, the software is easily adaptable to other problems requiring automated rapid responses of equipment to changes in the morphology of low contrast objects. (C) 2005 American Institute of Physics.