We demonstrate a complete nanotube electrophoresis system (nanotube radii in the range of 50 to 150 nm) based on lipid membranes, comprising DNA injection, single-molecule transport, and single-molecule detection. Using gel-capped electrodes, electrophoretic single-file transport of fluorescently labeled dsDNA molecules is observed inside nanotubes. The strong confinement to a channel of molecular dimensions ensures a detection efficiency close to unity and identification of DNA size from its linear relation to the integrated peak intensity. In addition to constituting a nanotechnological device for identification and quantification of single macromolecules or biopolymers, this system provides a method to study their conformational dynamics, reaction kinetics, and transport in cell-like environments.electrophoresis ͉ lipid ͉ conformation C ontrolled transport, interrogation, and manipulation of single molecules in integrated nanoscale devices would provide new tools for fundamental studies of molecular properties, development of ultrasensitive biochemical assays, and new models for studies of transport and reaction phenomena in confined biological systems. For example, it was recently shown that lipid bilayer nanotubes Ϸ50-200 nm in diameter are involved in mediated transport of water-soluble and membrane-bound components between cells (1). Such observations do not only raise important questions about transport mechanisms for macromolecules and organelles in spaces comparable to the size of the cargo itself, but also about how such strong confinement affects diffusion, conformation, and chemical reactions of enclosed molecules and particles (2-4). Furthermore, along with previous understanding of sorting and routing of individual molecules, e.g., in the Golgi-endoplasmic reticulum network (5, 6), these observations point out clearly that what has been an engineering dream for decades, i.e., to create manmade devices that can operate with single molecules in a controlled fashion, is a reality in biology and therefore can be a reality in the world of engineering provided that we procure sufficient knowledge and tools to emulate these systems. However, experimental systems for controlled confinement and transport of materials dissolved in fluids approaching the theoretical size limit, i.e., where the ratio of channel inner diameter and dimensions of the cargo are close to unity, have been difficult to make in combination with transport control. This difficulty is mainly because these systems have been fabricated by using solid-state materials and processing technologies used in the computer industry that are limited in terms of the smallest accessible length scales, topologies, materials properties, complexity of fabrication, and their necessary integration to large-scale instrumentation to drive fluid flow. Nonetheless, powerful micro͞nanofluidic protocols for polymer transport in solid-state nanochannels and pores (7-12) as well as in polydimethylsiloxane channels of a few micrometers in diameter (13, 14) have be...