A generalizable, tunable microfluidic platform for delivering fast temporally varying chemical signals to probe single-cell response dynamics.
A generalizable, tunable microfluidic platform for delivering fast temporally varying chemical signals to probe single-cell response dynamics.
复制标题
DOI：
10.1021/ac5019843
复制
发表时间：
2014-10-21
影响因子：
7.4
通讯作者：
Lu, Hang
中科院分区：
文献类型：
--
作者：
Chingozha, Loice;Zhan, Mei;Zhu, Cheng;Lu, Hang
了解生物系统如何将动态的、可溶的化学线索转化为生理过程，需要强大的实验工具来生成不同的时间化学模式。微流体的出现已经看到了快速流体交换平台的发展，使得细胞微环境的变化和精确的细胞处理变得容易。快速交换对于使系统暴露于时变信号是很重要的。然而，宏观流体流动与微观结构的直接耦合是潜在的问题，因为高剪切应力不可避免地会给感兴趣的生物系统增加混淆的机械扰动效应。在这里，我们设计了一种使用单片集成穿孔膜将快速和精确的宏观流动转换为微观流动的方法。我们集成了高密度细胞陷阱阵列，用于在流动条件下处理具有挑战性的非贴壁细胞和可溶性化学信号发生器模块。该平台能够在低剪应力下快速和可重复地切换刺激和缓冲，用于定量活的单细胞荧光研究。这种模块化设计可以方便地集成任何细胞处理芯片设计与任何化学输送模块。我们通过表征细胞在不同周期暴露于交替Ca2+波形的振荡响应的异质性来证明该装置的实用性。该平台能够在单细胞分辨率下分析细胞对化学扰动的反应，这对于理解信号转导途径是必要的。
Understanding how biological systems transduce dynamic, soluble chemical cues into physiological processes requires robust experimental tools for generating diverse temporal chemical patterns. The advent of microfluidics has seen the development of platforms for rapid fluid exchange allowing ease of changes in the cellular microenvironment and precise cell handling. Rapid exchange is important for exposing systems to temporally varying signals. However, direct coupling of macroscale fluid flow with microstructures is potentially problematic due to the high shear stresses that inevitably add confounding mechanical perturbation effects to the biological system of interest. Here, we have devised a method of translating fast and precise macroscale flows to microscale flows using a monolithically integrated perforated membrane. We integrated a high-density cell trap array for nonadherent cells that are challenging to handle under flow conditions with a soluble chemical signal generator module. The platform enables fast and repeatable switching of stimulus and buffer at low shear stresses for quantitative live, single-cell fluorescent studies. This modular design allows facile integration of any cell-handling chip design with any chemical delivery module. We demonstrate the utility of this device by characterizing heterogeneity of oscillatory response for cells exposed to alternating Ca2+ waveforms at various periodicities. This platform enables the analysis of cell responses to chemical perturbations at a single-cell resolution that is necessary in understanding signal transduction pathways.
登录
查看更多内容
影响因子：
56.9
作者：
Mettetal, Jerome T.;Muzzey, Dale;van Oudenaarden, Alexander
通讯作者：
van Oudenaarden, Alexander
DOI：
10.1073/pnas.87.7.2466
发表时间：
1990-04-01
影响因子：
11.1
作者：
THASTRUP, O;CULLEN, PJ;DAWSON, AP
通讯作者：
DAWSON, AP
影响因子：
6.1
作者：
Chung K;Zhan M;Srinivasan J;Sternberg PW;Gong E;Schroeder FC;Lu H
通讯作者：
Lu H
影响因子：
4.6
作者：
Arrol, H. P.;Church, L. D.;Young, S. P.
通讯作者：
Young, S. P.
影响因子：
3.4
作者：
Dalous, Jeremie;Burghardt, Emmanuel;Bretschneider, Till
通讯作者：
Bretschneider, Till