Microfluidic device for generating a stepwise concentration gradient on a microwell slide for cell analysis
2013 (English)In: Biomicrofluidics, ISSN 1932-1058, Vol. 7, no 6, 064115- p.Article in journal (Refereed) Published
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 μm-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.
Place, publisher, year, edition, pages
2013. Vol. 7, no 6, 064115- p.
Chemotaxis, Culture, Systems
Biomedical Laboratory Science/Technology
IdentifiersURN: urn:nbn:se:kth:diva-140190DOI: 10.1063/1.4846435ISI: 000329292200018ScopusID: 2-s2.0-84891443639OAI: oai:DiVA.org:kth-140190DiVA: diva2:688635
FunderVinnovaSwedish Foundation for Strategic Research Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 201401172014-01-172014-01-172014-01-30Bibliographically approved