We describe micromanipulation and microinjection procedures for the fabrication of soft-matter networks consisting of lipid bilayer nanotubes and surface-immobilized vesicles. these biomimetic membrane systems feature unique structural flexibility and expandability and, unlike solid-state microfluidic and nanofluidic devices prepared by top-down fabrication, they allow network designs with dynamic control over individual containers and interconnecting conduits. the fabrication is founded on self-assembly of phospholipid molecules, followed by micromanipulation operations, such as membrane electroporation and microinjection, to effect shape transformations of the membrane and create a series of interconnected compartments. size and geometry of the network can be chosen according to its desired function. Membrane composition is controlled mainly during the self-assembly step, whereas the interior contents of individual containers is defined through a sequence of microneedle injections. networks cannot be fabricated with other currently available methods of giant unilamellar vesicle preparation (large unilamellar vesicle fusion or electroformation). Described in detail are also three transport modes, which are suitable for moving water-soluble or membranebound small molecules, polymers, Dna, proteins and nanoparticles within the networks. the fabrication protocol requires ~90 min, provided all necessary preparations are made in advance. the transport studies require an additional 60-120 min, depending on the transport regime. 20,29 . Once a needle is inside a GUV, the bilayer tightly seals around the tip. The adhesion of the membrane is typically strong enough to allow the pulling of lipid material from the membrane, thereby instantly forming an open nanoconduit between the vesicle and the capillary. The flexible lipid nanotubes created in this way can be expanded at the needle-connected end by means of positive injection pressure, to an extent that a new vesicle is formed (Fig. 1, stage IV). Constant subsequent injection of fluid from the needle allows the 'daughter' vesicle to grow to a size appropriate for deposition onto the substrate (Fig. 1, stage V). Iteration of this pulling and vesiclegeneration process enables the building of whole networks of interconnected containers, in which size, geometry and individual contents can be controlled. If a new needle with different internal solution is used for the generation of each new vesicle, a network with differentiated contents can be created. The composition of the internal volume of each newly created vesicle is determined by the size ratio of mother and daughter vesicles 30 .As the nanoconduits are open on both ends and thus truly interconnect the individual containers, exchange of internal solution, and chemical compounds dissolved therein, is possible. Different regimes of exchange of matter have been investigated by us. In particular, the transport of small molecules and nanoparticles by diffusion [31][32][33] , by membrane tension-driven (Marangoni) flow 3...