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Gap junctional intercellular communication (GJIC) is critical to several physiological roles including cell proliferation,differentiation and apoptosis.l-3 Intercellular calcium wave (ICW) through gap junction may serve different functions.4 But paracrine like ATP-induced ATP release also results in ICW.5 So,a method for ruling out this possibility is in a great need.In this work,a microfluidic chip was designed and fabricated for researching on ATP-induced intercellular calcium signaling via gap junction in NIH-3T3 cells by two laminar flows.Cells were precisely moved into the channel and immobilized with high spatial resolution by using a negative pressure system.Therefore,the diffusion of ATP at the interface of two laminar flows would never touch the adjacent cells.ATP stimulated one part of a single adherent cell in one laminar flow and the intracellular Ca2+ concentration in adjacent cells in the other laminar flow changed following the first cell successfully.Even though ATP was released from the first cell,it would be washed downstream and could not enter into the other laminar flow.