Abstract: A microfluidic tissue dissociation and filtration device simultaneously filters large tissue fragments and dissociates smaller aggregates into single cells, thereby improving single cell yield and purity. The device includes an inlet coupled to a first microfluidic channel at an upstream location and a first outlet at a downstream location. A first filter membrane is interposed between the first microfluidic channel and a second microfluidic channel, wherein the second microfluidic channel is in fluidic communication with the first microfluidic channel via the first filter membrane. The first filter membrane operates under a tangential flow format. A second outlet is coupled to a downstream location of the second microfluidic channel and includes a second filter membrane interposed between the second outlet and the second microfluidic channel. The dual membrane device increased single cell numbers by at least 3-fold for different tissue types.