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Microfluidic device for generating a stepwise concentration gradient on a microwell slide for cell analysis

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Title: Microfluidic device for generating a stepwise concentration gradient on a microwell slide for cell analysis
Authors: Weibull, Emilie Browse this author
Matsui, Shunsuke Browse this author
Sakai, Manabu Browse this author
Svahn, Helene Andersson Browse this author
Ohashi, Toshiro Browse this author →KAKEN DB
Issue Date: Nov-2013
Publisher: Amer inst physics
Journal Title: Biomicrofluidics
Volume: 7
Issue: 6
Start Page: 064115-1
End Page: 064115-12
Publisher DOI: 10.1063/1.4846435
Abstract: Understanding biomolecular gradients and their role in biological processes is essential for fully comprehending the underlying mechanisms of cells in living tissue. Conventional in vitro gradient-generating methods are unpredictable and difficult to characterize, owing to temporal and spatial fluctuations. The field of microfluidics enables complex user-defined gradients to be generated based on a detailed understanding of fluidic behavior at the lm-scale. By using microfluidic gradients created by flow, it is possible to develop rapid and dynamic stepwise concentration gradients. However, cells exposed to stepwise gradients can be perturbed by signals from neighboring cells exposed to another concentration. Hence, there is a need for a device that generates a stepwise gradient at discrete and isolated locations. Here, we present a microfluidic device for generating a stepwise concentration gradient, which utilizes a microwell slide's pre-defined compartmentalized structure to physically separate different reagent concentrations. The gradient was generated due to flow resistance in the microchannel configuration of the device, which was designed using hydraulic analogy and theoretically verified by computational fluidic dynamics simulations. The device had two reagent channels and two dilutant channels, leading to eight chambers, each containing 4 microwells. A dose-dependency assay was performed using bovine aortic endothelial cells treated with saponin. High reproducibility between experiments was confirmed by evaluating the number of living cells in a live-dead assay. Our device generates a fully mixed fluid profile using a simple microchannel configuration and could be used in various gradient studies, e.g., screening for cytostatics or antibiotics. (C) 2013 AIP Publishing LLC.
Rights: Copyright 2013 American Institute of Physics. This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics. The following article appeared in Biomicrofluidics and may be found at
Type: article
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 大橋 俊朗

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