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The temperature-sensitive inverse macroporous membranes containing thermo-responsive poly (N-isopropylacrylamide) (PNIPAM) were prepared by a template of silica microspheres. The inverse membranes were characterized by SEM, FTIR and contact angle measurements. The PNIPAM containing inverse macroporous membranes had different pore diameters at different temperatures and could be used for separation. The conductivity was measured to characterize the temperature-responsive behavior and permeability of this inverse macroporous membranes. Due to variable pore sizes, the membranes exhibited specific temperature-responsive permeability. This strategy presented here is a simple, low-cost, and controllable alternative for fluidic applications, such as the separation of particles.