Experiments with gradients of soluble bioactive species have significantly advanced with microfluidic developments that enable cell observation and stringent control of environmental conditions. While some methodologies rely on flow to establish gradients, other opt for flow-free conditions, which is particularly beneficial for studying non-adherent and/or shear-sensitive cells. In flow-free devices, bioactive species diffuse either through resistive microchannels in “microchannel-based” devices, a porous membrane in “membrane-based” devices, or a hydrogel in “gel-based” devices. However, despite significant advancements over traditional methods such as “Boyden chambers”, these technologies have not widely disseminated in biological laboratories, arguably due to entrenched practices and the intricate skills required for conducting microfluidic assays. Here, we integrated Quake-type pneumatic microvalves in place of microgrooves, membranes, or gels, and developed devices with precise control over residual flow, establishment initial gradient, and long-term stability of gradients. The “Microvalve-based” approach enables the generation of the automatization of delicate microfluidic manipulations, which paves the way for routine applications of controlled and tunable flow-free gradients in academic laboratories and biomedical units.

DOI: https://doi.org/10.1039/D4LC00901K