Dynamic Generation of Concentration- and Temporal-Dependent Chemical Signals in an Integrated Microfluidic Device for Single-Cell Analysis

Analytical Chemistry
Alan M Gonzalez-SuarezJose L Garcia-Cordero

Abstract

Intracellular signaling pathways are affected by the temporal nature of external chemical signaling molecules such as neurotransmitters or hormones. Developing high-throughput technologies to mimic these time-varying chemical signals and to analyze the response of single cells would deepen our understanding of signaling networks. In this work, we introduce a microfluidic platform to stimulate hundreds of single cells with chemical waveforms of tunable frequency and amplitude. Our device produces a linear gradient of 9 concentrations that are delivered to an equal number of chambers, each containing 492 microwells, where individual cells are captured. The device can alternate between the different stimuli concentrations and a control buffer, with a maximum operating frequency of 33 mHz that can be adjusted from a computer. Fluorescent time-lapse microscopy enables to obtain hundreds of thousands of data points from one experiment. We characterized the gradient performance and stability by staining hundreds of cells with calcein AM. We also assessed the capacity of our device to introduce periodic chemical stimuli of different amplitudes and frequencies. To demonstrate our device performance, we studied the dynamics of intracellu...Continue Reading

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Citations

Aug 11, 2020·Clinical and Experimental Otorhinolaryngology·Se Hwan HwangAlexander Revzin
Mar 6, 2019·Analytical Chemistry·Diana F Cedillo-AlcantarAlexander Revzin
May 11, 2019·ACS Sensors·Xiaoyan LiuXingyu Jiang
Jan 14, 2020·ACS Sensors·Roberto Rodriguez-MoncayoJose Luis Garcia-Cordero
Oct 27, 2018·Analytical Chemistry·Pieter E OomenAndrew G Ewing

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