We report on a fundamental technological advance for multilayer polydimethylsiloxane (PDMS) microfluidics. Vertical passages (vias), connecting channels located in different layers, are fabricated monolithically, in parallel, by simple and easy means. The resulting 3D connectivity greatly expands the potential complexity of microfluidic architecture. We apply the vias to printing nested bioarrays and building autoregulatory devices. A current source is demonstrated, while a diode and a rectifier are derived; all are building blocks for analog circuitry in Newtonian fluids. We also describe microfluidic septa and their applications. Vias lay the foundation for a new generation of microfluidic devices.ver the decade of its existence, polydimethylsiloxane (PDMS) microfluidics has progressed from the plain microchannel (1) through pneumatic valves and pumps (2, 3) to an impressive set of specialized components organized by the thousands in multilayer large-scale-integration chips (4). These devices have become the hydraulic elastomeric embodiment of Richard Feynman's dreams of infinitesimal machines (5, 6). The now established technology (7) has found successful applications in protein crystallization (8), DNA sequencing (9), nanoliter PCR (10), cell sorting and cytometry (11), nucleic acids extraction and purification (12), immunoassays (13,14), cell studies (15-18), and chemical synthesis (19), while also serving as the fluid-handling component in emerging integrated microelectromechanical devices (MEMS) (20).The energetic pursuit of applications, however, has resulted in a premature attention shift away from fundamental microfluidics. Here we report on a fundamental technological advance that allows a simple and easy access to a large increase in the architectural complexity of microfluidic devices, as well as opens new possibilities for technical developments and consequent applications. We dubbed the previously undescribed device ''via,'' in reference to its analog in modern semiconductor electronics.Vias are vertical micropassages that connect channels fabricated in different layers of the same PDMS multilayer chip. The functional result is 3D channels that lift the restrictions of the traditional architecture wherein channels could not leave their layer and two channels within the same layer could not cross without mixing. These restrictions did not prevent the emergence of expansive architectures (4), because the particular applications were shrewdly chosen to involve large-scale parallelization of simple identical operations, thereby requiring few controls and maximizing device density. However, as the field moves to functionally complex heterogeneous devices integrated on the same chip, laying out the respective circuitry would inevitably necessitate convenient, simple, and reliable vertical connectivity just as it did in the semiconductor industry. Microfluidic vias provide that 3D connectivity, lift the above architectural restrictions, and contribute morphological and functional capabilities.The pursuit...