Fluidic transport through nanochannels offers new opportunities to probe fundamental nanoscale transport phenomena and to develop tools for manipulating DNA, proteins, small molecules and nanoparticles. The small size of nanofabricated devices and the accompanying increase in the effect of surface forces, however, pose challenges in designing and fabricating flexible nanofluidic systems that can dynamically adjust their transport characteristics according to the handling needs of various molecules and nanoparticles. Here, we describe the use of nanoscale fracturing of oxidized poly(dimethylsiloxane) to conveniently fabricate nanofluidic systems with arrays of nanochannels that can actively manipulate nanofluidic transport through dynamic modulation of the channel cross-section. We present the design parameters for engineering material properties and channel geometry to achieve reversible nanochannel deformation using remarkably small forces. We demonstrate the versatility of the elastomeric nanochannels through tuneable sieving and trapping of nanoparticles, dynamic manipulation of the conformation of single DNA molecules and in situ photofabrication of movable polymeric nanostructures.
Computer-controlled microfluidics would advance many types of cellular assays and microscale tissue engineering studies wherever spatiotemporal changes in fluidics need to be defined. However, this goal has been elusive because of the limited availability of integrated, programmable pumps and valves. This paper demonstrates how a refreshable Braille display, with its grid of 320 vertically moving pins, can power integrated pumps and valves through localized deformations of channel networks within elastic silicone rubber. The resulting computerized fluidic control is able to switch among: (i) rapid and efficient mixing between streams, (ii) multiple laminar flows with minimal mixing between streams, and (iii) segmented plug-flow of immiscible fluids within the same channel architecture. The same control method is used to precisely seed cells, compartmentalize them into distinct subpopulations through channel reconfiguration, and culture each cell subpopulation for up to 3 weeks under perfusion. These reliable microscale cell cultures showed gradients of cellular behavior from C2C12 myoblasts along channel lengths, as well as differences in cell density of undifferentiated myoblasts and differentiation patterns, both programmable through different flow rates of serumcontaining media. This technology will allow future microscale tissue or cell studies to be more accessible, especially for highthroughput, complex, and long-term experiments. The microfluidic actuation method described is versatile and computer programmable, yet simple, well packaged, and portable enough for personal use.pump ͉ valve ͉ bioreactor ͉ mixer ͉ perfusion A dvanced microfluidic cellular assays (1-6) and microscale tissue engineering studies (7-10) would benefit from robust and convenient methods to computer-control accurate spatiotemporal patterns of microfluidic flows in arrays of fluidic networks. In the past, fluidic control included syringe pumps (11), hydrogel valves (12), gravity-driven pumps (13), evaporation-based pumps (14, 15), acoustic pumps (16), gas-generationbased pumps (17, 18), and centrifugal force in CD chips (19). Electrokinetic flow (20-22), thermopneumatic (23, 24), pneumatic͞hydraulic (25), or mechanical (26, 27) valves and pumps have a high degree of control, but require external connection lines to larger equipment for actuation. A few fully integrated and self-packaged systems (23, 28) have been developed, but lack the reconfigurability inherent with numerous active valves and pumps.Here, we report a method to precisely control fluid flow inside elastomeric capillary networks by using multiple (tens to hundreds) computer-controlled, piezoelectric, movable pins. These pins are positioned as a grid on a refreshable Braille display (e.g., F. H. Papenmeier, Schwerte, Germany), which is a tactile device used by the visually impaired to read computer text. Each pin can act as a valve and be shifted upward to push against channels contained in silicone rubber and completely shut the channel. Three sequential valves of this...
This paper describes a self-contained integrated microfluidic system that can separate motile sperm from small samples that are difficult to handle using conventional sperm-sorting techniques. The device isolates motile sperm from nonmotile sperm and other cellular debris, based on the ability of motile sperm to cross streamlines in a laminar fluid stream. The device is small, simple, and disposable yet is an integrated system complete with sample inlets, outlets, sorting channel, and a novel passively driven pumping system that provides a steady flow of liquid; it requires no external power source or controls. The device fulfills a need in clinical settings where small amounts of sperm need to be sorted. It also opens the way for convenient bioassays based on sperm motility including at-home motile sperm tests.
The interface between extracellular matrices and cells is a dynamic environment that is crucial for regulating important cellular processes such as signal transduction, growth, differentiation, motility and apoptosis. In vitro cellular studies and the development of new biomaterials would benefit from matrices that allow reversible modulation of the cell adhesive signals at a scale that is commensurate with individual adhesion complexes. Here, we describe the fabrication of substrates containing arrays of cracks in which cell-adhesive proteins are selectively adsorbed. The widths of the cracks (120-3,200 nm) are similar in size to individual adhesion complexes (typically 500-3,000 nm) and can be modulated by adjusting the mechanical strain applied to the substrate. Morphology of cells can be reversibly manipulated multiple times through in situ adjustment of crack widths and hence the amount of the cell-adhesive proteins accessible to the cell. These substrates provide a new tool for assessing cellular responses associated with exposure to matrix proteins.
Techniques for introducing foreign molecules and materials into living cells are of great value in cell biology research. A major barrier for intracellular delivery is to cross the cell membrane. Here we demonstrate a novel platform utilizing diamond nanoneedle arrays to facilitate efficient vector-free cytosolic delivery. Using our technique, cellular membrane is deformed by an array of nanoneedles with a force on the order of a few nanonewtons. We show that this technique is applicable to deliver a broad range of molecules and materials into different types of cells, including primary neurons in adherent culture. Especially, for delivering plasmid DNAs into neurons, our technique produces at least eightfold improvement (B45% versus B1-5%) in transfection efficiency with a dramatically shorter experimental protocol, when compared with the commonly used lipofection approach. It is anticipated that our technique will greatly benefit basic research in cell biology and also a wide variety of clinical applications.
Theranostic nanomedicine is capable of diagnosis, therapy, and monitoring the delivery and distribution of drug molecules and has received growing interest. Herein, a self-monitored and self-delivered photosensitizer-doped FRET nanoparticle (NP) drug delivery system (DDS) is designed for this purpose. During preparation, a donor/acceptor pair of perylene and 5,10,15,20-tetro (4-pyridyl) porphyrin (H2TPyP) is co-doped into a chemotherapeutic anticancer drug curcumin (Cur) matrix. In the system, Cur works as a chemotherapeutic agent. In the meantime, the green fluorescence of Cur molecules is quenched (OFF) in the form of NPs and can be subsequently recovered (ON) upon release in tumor cells, which enables additional imaging and real-time self-monitoring capabilities. H2TPyP is employed as a photodynamic therapeutic drug, but it also emits efficient NIR fluorescence for diagnosis via FRET from perylene. By exploiting the emission characteristics of these two emitters, the combinatorial drugs provide a real-time dual-fluorescent imaging/tracking system in vitro and in vivo, and this has not been reported before in self-delivered DDS which simultaneously shows a high drug loading capacity (77.6%Cur). Overall, our carrier-free DDS is able to achieve chemotherapy (Cur), photodynamic therapy (H2TPyP), and real-time self-monitoring of the release and distribution of the nanomedicine (Cur and H2TPyP). More importantly, the as-prepared NPs show high cancer therapeutic efficiency both in vitro and in vivo. We expect that the present real-time self-monitored and self-delivered DDS with multiple-therapeutic and multiple-fluorescent ability will have broad applications in future cancer therapy.
Improved polyvinylpyrrolidone (PVP) microneedle arrays can be fabricated by adding cyclodextrin (CD) to form PVP–CD inclusion complexes.
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