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Parallel Multi-Time Point Cell Stimulus and Lysis in a Microfluidic Device Using Chaotic Mixing and Pressure Resistance

Journal article published in 2008 by Alison Hirsch, Catherine Rivet, Boyang Zhang, Melissa Kemp, Hang Lu ORCID
This paper is available in a repository.
This paper is available in a repository.

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Abstract

A high-throughput two-module device was designed to simulatiously obtain 8 time points of cell activation through stimulation with 30s resolution rapidly and ac-curately. Rapid mixing and even splitting of reagents into each time-point channel are necessary features of this device. The microfluidic platform allows uniform sample handeling with superior temporal resolution compared to conventional meth-ods. INTRODUCTION Understanding the dynamics of cell signaling networks is important for many biological applications. To build a complete computational model of the dynamic pathway a large data set is necessary. Gene expression and protein activity at vari-ous time points during stimulation with an external signal must be known. Many important protein activation events occur within minutes after stimulation [1, 2]. However, with conventional multi-well plate assays, it is difficult to achieve ade-quate resolution in the appropriate time scales. Microfluidics is a capable alterna-tive, providing uniformity in sample handling to reduce error between experiments [3]. We present here a device for multi time-point lymphocyte stimulation and lysis for downstream analysis of protein activation. Previously El-Ali et al. developed a device achieving cell stimulus and lysis on a microfluidic chip using segmented gas-liquid flow for rapid mixing [4]. This device was the first to demonstrate on-chip multiple step manipulation of cells with fast mixing although with only one time point per experiment and large shear at the gas-liquid interface. In comparison, our device is capable of 8 time points with controlled rapid mixing, precise timed stimu-lation, and rapid lysis. EXPERIMENTAL This two-module device is molded in PDMS from a two-layer SU-8 master. During operation (Fig. 1), a syringe pump drives the flow to only 3 inlets, cells and stimulus are mixed and split into 8 equal streams in Module 1. The majority of the incubation time occurs in the tubing leading to Module 2. There the reaction is quenched and cells are burst with lysis buffer to extract intracellular components.