Parallel multi-time point cell stimulation and lysis on-chip for studying early signaling events in T cell activation.
Parallel multi-time point cell stimulation and lysis on-chip for studying early signaling events in T cell activation.
复制标题
DOI：
10.1039/b810896j
复制
发表时间：
2009-02-21
期刊：
影响因子：
6.1
通讯作者：
Lu H
中科院分区：
文献类型：
--
作者：
Hirsch AM;Rivet CA;Zhang B;Kemp ML;Lu H
Dynamics of complex signaling networks are important to many biological problems. Quantitative data at early time points after cellular stimulation are necessary for accurate model generation. However, the large amount of data needed is often extremely time-consuming and expensive to acquire with conventional methods. We present a two-module microfluidic platform for simultaneous multi-time point stimulation and lysis of T-Cells for early time point signaling activation with a resolution down to 20 seconds using only small amounts of cells and reagents. The key design features are rapid mixing of reagents and uniform splitting into 8 channels for simultaneous collection of multi-time point data. Chaotic mixing was investigated via computational fluid dynamic modeling, and was used to achieve rapid and complete mixing. This modular device is flexible - with easy adjustment of the setup, a wide range of time points can be achieved. We show that treatment in the device does not elicit adverse cellular stress in Jurkat cells. The activation of 6 important proteins in the signaling cascade was quantified upon stimulation with a soluble form of α-CD3. The dynamics from device and conventional methods are similar, but the micro device exhibits significantly less error between experiments. We envision this high-throughput format to enable simple and fast generation of large sets of quantitative data, with consistent sample handling, for many complex biological systems.
登录
查看更多内容
影响因子：
4.4
作者：
Kemp, Melissa L.;Wille, Lucia;Lauffenburger, Douglas A.
通讯作者：
Lauffenburger, Douglas A.
影响因子：
3.9
作者：
Kohrt, HE;Shu, CT;Lee, PP
通讯作者：
Lee, PP
影响因子：
64.8
作者：
Reya, T;Clevers, H
通讯作者：
Clevers, H
影响因子：
6.1
作者：
King, Kevin R.;Wang, Sihong;Toner, Mehmet
通讯作者：
Toner, Mehmet
DOI：
10.1073/pnas.0607254104
发表时间：
2007-03-27
影响因子：
11.1
作者：
Herr, Amy E.;Hatch, Anson V.;Singh, Anup K.
通讯作者：
Singh, Anup K.