The concentration control of reagents is an important factor in microfluidic devices for cell cultivation and chemical mixing, but it is difficult to realize owing to the characteristics of microfluidic devices. We developed a microfluidic device that can generate concentration gradients among multiple main chambers. Multiple main chambers are connected in parallel to the body channel via the neck channel. The main chamber is subjected to a volume change through a driving chamber that surrounds the main chamber, and agitation is performed on the basis of the inequality of flow caused by expansion or contraction. The neck channel is connected tangentially to the main chamber. When the main chamber expands or contracts, the flow in the main chamber is unequal, and a net vortex is generated. The liquid moving back and forth in the neck channel gradually absorbs the liquid in the body channel into the main chamber. As the concentration in the main chamber changes depending on the pressure applied to the driving chamber, we generated a concentration gradient by arranging chambers along the pressure gradient. This allowed for us to create an environment with different concentrations on a single microchip, which is expected to improve observation efficiency and save space.