A robust diffusion-based gradient generator for dynamic cell assays
A robust diffusion-based gradient generator for dynamic cell assays
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DOI：
10.1039/c1lc20829b
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发表时间：
2012-01-01
期刊：
影响因子：
6.1
通讯作者：
Locascio, Laurie E.
中科院分区：
文献类型：
--
作者：
Atencia, Javier;Cooksey, Gregory A.;Locascio, Laurie E.
这份手稿描述了一种新的方法来产生纯扩散的化学梯度，可以及时修改。该装置设计简单，易于使用，这使得它适合研究涉及静态或动态化学梯度的生物过程，如趋化性。我们描述了无对流梯度发生器的理论基础，说明了实现该理论的设计，并提出了一个协议，以调整多层双面胶带和层压板来制造设备。使用该装置，在48小时实验中，将哺乳动物细胞群体暴露于浓度梯度内的不同浓度的毒素。通过循环梯度开和关动态探测细胞，并使用延时荧光显微镜监测细胞反应。实验和结果说明了涉及动态细胞行为的应用程序的类型，可以使用这种类型的梯度生成器作为目标。
This manuscript describes a new method to generate purely diffusive chemical gradients that can be modified in time. The device is simple in its design and easy to use, which makes it amenable to study biological processes that involve static or dynamic chemical gradients such as chemotaxis. We describe the theory underlying the convection-free gradient generator, illustrate the design to implement the theory, and present a protocol to align multiple layers of double sided tape and laminates to fabricate the device. Using this device, a population of mammalian cells was exposed to different concentrations of a toxin within a concentration gradient in a 48 h experiment. Cells were probed dynamically by cycling the gradient on and off, and cell response was monitored using time-lapse fluorescence microscopy. The experiment and results illustrate the type of applications involving dynamic cell behavior that can be targeted with this type of gradient generator.