Exosomes, the smallest sized extracellular vesicles (30 ~ 150 nm) packaged with lipids, proteins, functional messenger RNAs and microRNAs, and double-stranded DNA from their cells of origin, have emerged as key players in intercellular communication. Their presence in bodily fluids, where they protect their cargo from degradation, makes them attractive candidates for clinical application as innovative diagnostic and therapeutic tools. But routine isolation and analysis of high purity exosomes in clinical settings is challenging, with conventional methods facing a number of drawbacks including low yield and/or purity, long processing times, high cost, and difficulties in standardization. Here we review a promising solution, microfluidic-based technologies that have incorporated a host of separation and sensing capabilities for exosome isolation, detection, and analysis, with emphasis on point of care and clinical applications. These new capabilities promise to advance fundamental research while paving the way toward routine exosome-based liquid biopsy for personalized medicine.
The ability to mix liquids in microchannel networks is fundamentally important in the design of nearly every miniaturized chemical and biochemical analysis system. Here, we show that enhanced micromixing can be achieved in topologically simple and easily fabricated planar 2D microchannels by simply introducing curvature and changes in width in a prescribed manner. This goal is accomplished by harnessing a synergistic combination of (i) Dean vortices that arise in the vertical plane of curved channels as a consequence of an interplay between inertial, centrifugal, and viscous effects, and (ii) expansion vortices that arise in the horizontal plane due to an abrupt increase in a conduit's cross-sectional area. We characterize these effects by using confocal microscopy of aqueous fluorescent dye streams and by observing binding interactions between an intercalating dye and double-stranded DNA. These mixing approaches are versatile and scalable and can be straightforwardly integrated as generic components in a variety of lab-on-a-chip systems.Dean flow ͉ expansion vortex ͉ microfluidics ͉ lab on a chip A lthough microf luidic mixing is a key process in a host of miniaturized analysis systems (1-7), it continues to pose challenges owing to constraints associated with operating in an unfavorable laminar f low regime dominated by molecular diffusion and characterized by a combination of low Reynolds numbers (Re ϭ Vd͞v Ͻ Ͻ 100, where V is the f low velocity, d is a length scale associated with the channel diameter, and v is the f luid kinematic viscosity) and high Péclet numbers (Pe ϭ Vd͞D Ͼ 100, where D is the molecular diffusivity). The relatively large discrepancy between convective and diffusive timescales implies that in a straight smooth-walled microchannel, the downstream distances over which liquids must travel to become fully intermixed (⌬y m ϳ Vd 2 ͞D ϭ Pe ϫ d) can be on the order of several centimeters. These mixing lengths are generally prohibitively long and often negate many of the benefits of miniaturization.A wide variety of micromixing approaches have been explored (8, 9), most of which can be broadly classified as either ''active'' (involving input of external energy) or ''passive'' (harnessing the inherent hydrodynamic structure of specific flow fields to mix fluids in the absence of external forces). Passive designs are often desirable in applications involving sensitive species (e.g., biological samples) because they do not impose strong mechanical, electrical, or thermal agitation. Examples of passive micromixing approaches that have been widely investigated include the following: (i) ''split-and-recombine'' strategies where the streams to be mixed are divided or split into multiple channels and redirected along trajectories that allow them to be subsequently reassembled as alternating lamellae yielding exponential reductions in interspecies diffusion length and time scales (4, 10-12); and (ii) ''chaotic'' strategies where transverse flows are passively generated that continuously expand interfacial...
An integrated microfluidic device capable of performing a variety of genetic assays has been developed as a step towards building systems for widespread dissemination. The device integrates fluidic and thermal components such as heaters, temperature sensors, and addressable valves to control two nanoliter reactors in series followed by an electrophoretic separation. This combination of components is suitable for a variety of genetic analyses. As an example, we have successfully identified sequence-specific hemagglutinin A subtype for the A/LA/1/87 strain of influenza virus. The device uses a compact design and mass production technologies, making it an attractive platform for a variety of widely disseminated applications.
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