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Acoustofluidic devices for manipulating microparticles in fluids are appealing for biological sample processing due to their gentle and high-speed capability of sorting cell-scale objects. Such devices are generally limited to moving particles toward locations at integer fractions of the fluid channel width (1/2, 1/4, 1/6, etc.). In this work, we introduce a unique approach to acoustophoretic device design that overcomes this constraint, allowing us to design the particle focusing location anywhere within the microchannel. This is achieved by fabricating a second fluid channel in parallel with the sample channel, separated from it by a thin silicon wall. The fluids in both channels participate to create the ultrasound resonance, while only one channel processes the sample, thus de-coupling the fluidic and acoustic boundaries. The wall placement and the relative widths of the adjacent channels define the particle focusing location. We investigate the operating characteristics of a range of these devices to determine the configurations that enable effective particle focusing and separation. The results show that a sufficiently thin wall negligibly affects focusing efficiency and location compared to a single channel without a wall, validating the success of this design approach without compromising separation performance. Using these principles to design and fabricate an optimized device configuration, we demonstrate high-efficiency focusing of microspheres, as well as separation of cell-free viruses from mammalian cells. These "transparent wall" acoustic devices are capable of over 90% extraction efficiency with 10 µm microspheres at 450 µL min(-1), and of separating cells (98% purity), from viral particles (70% purity) at 100 µL min(-1).
Asunto(s)
Acústica , Virus del Dengue/aislamiento & purificación , Técnicas Analíticas Microfluídicas/métodos , Tamaño de la Partícula , Animales , Bioingeniería/métodos , Chlorocebus aethiops , Técnicas Analíticas Microfluídicas/normas , Microesferas , Células VeroRESUMEN
Digital PCR enables the absolute quantitation of nucleic acids in a sample. The lack of scalable and practical technologies for digital PCR implementation has hampered the widespread adoption of this inherently powerful technique. Here we describe a high-throughput droplet digital PCR (ddPCR) system that enables processing of ~2 million PCR reactions using conventional TaqMan assays with a 96-well plate workflow. Three applications demonstrate that the massive partitioning afforded by our ddPCR system provides orders of magnitude more precision and sensitivity than real-time PCR. First, we show the accurate measurement of germline copy number variation. Second, for rare alleles, we show sensitive detection of mutant DNA in a 100,000-fold excess of wildtype background. Third, we demonstrate absolute quantitation of circulating fetal and maternal DNA from cell-free plasma. We anticipate this ddPCR system will allow researchers to explore complex genetic landscapes, discover and validate new disease associations, and define a new era of molecular diagnostics.
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ADN/genética , Dosificación de Gen/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Reacción en Cadena de la Polimerasa , HumanosRESUMEN
We present a theoretical and experimental study of the role of hydrodynamic interactions on the motion and dispersion of metal rodlike particles in the presence of an externally applied electric field. In these systems, the electric field polarizes the particles and induces an electroosmotic flow relative to the surface of each particle. The simulations include the effect of the gravitational body force, buoyancy, far-field hydrodynamic interactions, and near-field lubrication forces. The particles in the simulations and experiments were observed to experience repeated pairing interactions in which they come together axially with their ends approaching each other, slide past one another until their centers approach, and then push apart. These interactions were confirmed in measurements of particle orientations and velocities, pair distribution functions, and net dispersion of the suspension. For large electric fields, the pair distribution functions show accumulation and depletion regions consistent with many pairing events. For particle concentrations of 10;{8}particles/mL and higher, dispersion within the suspension dramatically increases with increased field strength.
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This article presents analysis and optimization of a microfluidic particle filter that uses acoustic radiation forces to remove particles larger than a selected size by adjusting the driving conditions of the piezoelectric transducer (PZT). Operationally, the acoustic filter concentrates microparticles to the center of the microchannel, minimizing undesirable particle adsorption to the microchannel walls. Finite element models predict the complex two-dimensional acoustic radiation force field perpendicular to the flow direction in microfluidic devices. We compare these results with experimental parametric studies including variations of the PZT driving frequencies and voltages as well as various particle sizes (0.5-5.0 microm in diameter). These results provide insight into the optimal operating conditions and show the efficacy of our device as a filter with an adjustable effective pore size. We demonstrate the separation of Saccharomyces cerevisiae from MS2 bacteriophage using our acoustic device. With optimized design of our microfluidic flow system, we achieved yields of greater than 90% for the MS2 with greater than 80% removal of the S. cerevisiae in this continuous-flow sample preparation device.
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Acústica , Métodos Analíticos de la Preparación de la Muestra/métodos , Técnicas Analíticas Microfluídicas/métodos , Automatización , Levivirus/aislamiento & purificación , Porosidad , Saccharomyces cerevisiae/citología , Saccharomyces cerevisiae/aislamiento & purificación , Factores de TiempoRESUMEN
Analytical models are developed for the translation and rotation of metallic rods in a uniform electric field. The limits of thin and thick electric double layers are considered. These models include the effect of stripes of different metals along the length of the particle. Modeling results are compared to experimental measurements for metallic rods. Experiments demonstrate the increased alignment of particles with increasing field strength and the increase in degree of alignment of thin versus thick electric double layers. The metal rods polarize in the applied field and align parallel to its direction due to torques on the polarized charge. The torque due to polarization has a second-order dependence on the electric field strength. The particles are also shown to have an additional alignment torque component due to nonuniform densities along their length. The orientation distributions of dilute suspensions of particles are also shown to agree well with results predicted by a rotational convective-diffusion equation.
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BACKGROUND: international estimates suggest that caesarean section on maternal request range from 4% to 18% of all caesarean section. An increasing number of surveys have investigated women's reasons for a caesarean section in the absence of a medical indication but few studies have solely studied first-time mothers motivation for this request. OBJECTIVE: to describe the underlying reasons for the desire for a caesarean section in the absence of medical indication in pregnant first-time mothers. METHOD: a qualitative descriptive study, with content analysis of interviews with 12 first-time mothers. FINDINGS: the overarching theme formulated to illustrate the central interpreted meaning of the underlying desire for a planned caesarean section was based on deeply rooted emotions'. Four categories were identified as related to the request for a caesarean section on maternal request. The categories was identified as 'always knowing that there are no other options than a caesarean section', 'caesarean section as a more controlled and safe way of having a baby', own negative experiences of health care and having problems dealing with other people's reaction about their mode of delivery. CONCLUSION: the results show that for these first-time mothers deeply rooted emotions described as stronger than fear of birth were behind their wish for a planned caesarean section.
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Cesárea/psicología , Emociones , Paridad , Prioridad del Paciente , Mujeres Embarazadas/psicología , Adulto , Cesárea/estadística & datos numéricos , Toma de Decisiones , Femenino , Humanos , Participación del Paciente , Prioridad del Paciente/psicología , Prioridad del Paciente/estadística & datos numéricos , Embarazo , Investigación Cualitativa , SueciaRESUMEN
PURPOSE: The objective of the study was to compare patterns of survival 2001-2004 in prostate cancer patients from England, Norway and Sweden in relation to age and period of follow-up. SUBJECTS AND METHODS: Excess mortality in men with prostate cancer was estimated using nation-wide cancer register data using a period approach for relative survival. 179,112 men in England, 23,192 in Norway and 59,697 in Sweden were included. RESULTS: In all age groups, England had the lowest survival, particularly so among men aged 80+. Overall age-standardised five-year survival was 76.4%, 80.3% and 83.0% for England, Norway and Sweden, respectively. The majority of the excess deaths in England were confined to the first year of follow-up. CONCLUSION: The results indicate that a small but important group of older patients present at a late stage and succumb early to their cancers, possibly in combination with severe comorbidity, and this situation is more common in England than in Norway or Sweden.
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Neoplasias de la Próstata/mortalidad , Anciano , Anciano de 80 o más Años , Inglaterra/epidemiología , Humanos , Masculino , Tamizaje Masivo , Noruega/epidemiología , Neoplasias de la Próstata/epidemiología , Práctica de Salud Pública , Tasa de Supervivencia , Suecia/epidemiologíaRESUMEN
Metallic nanoparticles suspended in aqueous solutions and functionalized with chemical and biological surface coatings are important elements in basic and applied nanoscience research. Many applications require an understanding of the electrokinetic or colloidal properties of such particles. We describe the results of experiments to measure the zeta potential of metallic nanorod particles in aqueous saline solutions, including the effects of pH, ionic strength, metallic composition, and surface functionalization state. Particle substrates tested include gold, silver, and palladium monometallic particles as well as gold/silver bimetallic particles. Surface functionalization conditions included 11-mercaptoundecanoic acid (MUA), mercaptoethanol (ME), and mercaptoethanesulfonic acid (MESA) self-assembled monolayers (SAMs), as well as MUA layers subsequently derivatized with proteins. For comparison, we present zeta potential data for typical charge-stabilized polystyrene particles. We compare experimental zeta potential data with theoretically predicted values for SAM-coated and bimetallic particles. The results of these studies are useful in predicting and controlling the aggregation, adhesion, and transport of functionalized metallic nanoparticles within microfluidic devices and other systems.
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Electroforesis/métodos , Nanopartículas del Metal/química , Electricidad , Ácidos Grasos/química , Oro/química , Modelos Teóricos , Compuestos de Sulfhidrilo/químicaRESUMEN
We modeled temporal trends in the 1- and 5-year survival of 32 499 patients with adenocarcinoma and squamous cell carcinoma of the lung in the Swedish Cancer Register between 1961 and 2000. The 1-year relative survival for adenocarcinoma improved from 37% for patients diagnosed 1961-1965 to 45% for those diagnosed 1996-2000 and from 39 to 45% for squamous cell carcinoma. The adjusted excess mortality ratios for the period 1996-2000 compared with 1961-1965 were 0.80 for adenocarcinoma and 0.81 for squamous cell carcinoma. Thus, a previous report in a Dutch study of a relatively worsening prognosis for adenocarcinoma over time could not be confirmed.
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Adenocarcinoma/mortalidad , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Carcinoma de Células Escamosas/mortalidad , Neoplasias Pulmonares/mortalidad , Adenocarcinoma/etiología , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma de Pulmón de Células no Pequeñas/etiología , Carcinoma de Células Escamosas/etiología , Femenino , Humanos , Neoplasias Pulmonares/etiología , Masculino , Persona de Mediana Edad , Fumar/efectos adversos , Suecia/epidemiología , Factores de TiempoRESUMEN
The first lab-on-chip system for picoliter droplet generation and PCR amplification with real-time fluorescence detection has performed PCR in isolated droplets at volumes 106 smaller than commercial real-time PCR instruments. The system utilized a shearing T-junction in a silicon device to generate a stream of monodisperse picoliter droplets that were isolated from the microfluidic channel walls and each other by the oil-phase carrier. An off-chip valving system stopped the droplets on-chip, allowing them to be thermally cycled through the PCR protocol without droplet motion. With this system, a 10-pL droplet, encapsulating less than one copy of viral genomic DNA through Poisson statistics, showed real-time PCR amplification curves with a cycle threshold of approximately 18, 20 cycles earlier than commercial instruments. This combination of the established real-time PCR assay with digital microfluidics is ideal for isolating single-copy nucleic acids in a complex environment.