RESUMEN
In this work, we present the results of a comparison of simple artificial neural network (FFNN) designs intended to identify infant bottle-feeding events and appropriate feeding volume recording intervals using accelerometer data recorded from a custom designed "Smart Bottle" system. To properly identify and distinguish these events with an accuracy of 99.8%, while accommodating the constraints of the deployment environment, two concurrent FFNNs were implemented.
Asunto(s)
Alimentación con Biberón , Redes Neurales de la Computación , Humanos , LactanteRESUMEN
The applications of dielectrophoretic (DEP) techniques for the manipulation of cells in a label-free fashion within microfluidic systems continue to grow. However, a limited number of methods exist for making highly sensitive separations that can isolate subtle phenotypic differences within a population of cells. This paper explores efforts to leverage that most compelling aspect of DEP-an actuation force that depends on particle electrical properties-in the background of phenotypic variations in cell size. Several promising approaches, centering around the application of multiple electric fields with spatially mapped magnitude and/or frequencies, are expanding the capability of DEP cell separation.
RESUMEN
The capture of circulating tumor cells (CTCs) from cancer patient blood enables early clinical assessment as well as genetic and pharmacological evaluation of cancer and metastasis. Although there have been many microfluidic immunocapture and electrokinetic techniques developed for isolating rare cancer cells, these techniques are often limited by a capture performance tradeoff between high efficiency and high purity. We present the characterization of shear-dependent cancer cell capture in a novel hybrid DEP-immunocapture system consisting of interdigitated electrodes fabricated in a Hele-Shaw flow cell that was functionalized with a monoclonal antibody, J591, which is highly specific to prostate-specific membrane antigen expressing prostate cancer cells. We measured the positive and negative DEP response of a prostate cancer cell line, LNCaP, as a function of applied electric field frequency, and showed that DEP can control capture performance by promoting or preventing cell interactions with immunocapture surfaces, depending on the sign and magnitude of the applied DEP force, as well as on the local shear stress experienced by cells flowing in the device. This work demonstrates that DEP and immunocapture techniques can work synergistically to improve cell capture performance, and it will aid in the design of future hybrid DEP-immunocapture systems for high-efficiency CTC capture with enhanced purity.
Asunto(s)
Separación Celular/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Células Neoplásicas Circulantes/patología , Neoplasias de la Próstata/patología , Anticuerpos Inmovilizados/química , Anticuerpos Monoclonales/química , Línea Celular Tumoral , Electrodos , Diseño de Equipo , Humanos , Masculino , Neoplasias de la Próstata/sangreRESUMEN
This article reviews existing methods for the isolation, fractionation, or capture of rare cells in microfluidic devices. Rare cell capture devices face the challenge of maintaining the efficiency standard of traditional bulk separation methods such as flow cytometers and immunomagnetic separators while requiring very high purity of the target cell population, which is typically already at very low starting concentrations. Two major classifications of rare cell capture approaches are covered: (1) non-electrokinetic methods (e.g., immobilization via antibody or aptamer chemistry, size-based sorting, and sheath flow and streamline sorting) are discussed for applications using blood cells, cancer cells, and other mammalian cells, and (2) electrokinetic (primarily dielectrophoretic) methods using both electrode-based and insulative geometries are presented with a view towards pathogen detection, blood fractionation, and cancer cell isolation. The included methods were evaluated based on performance criteria including cell type modeled and used, number of steps/stages, cell viability, and enrichment, efficiency, and/or purity. Major areas for improvement are increasing viability and capture efficiency/purity of directly processed biological samples, as a majority of current studies only process spiked cell lines or pre-diluted/lysed samples. Despite these current challenges, multiple advances have been made in the development of devices for rare cell capture and the subsequent elucidation of new biological phenomena; this article serves to highlight this progress as well as the electrokinetic and non-electrokinetic methods that can potentially be combined to improve performance in future studies.
RESUMEN
We report the positive dielectrophoretic (pDEP) characterization of wild-type and ethambutol-treated Mycobacterium smegmatis populations via automated pDEP cell trapping experiments. The automated technique was validated by measurements of carboxylate-modified polystyrene microspheres and Escherichia coli . The characterization of M. smegmatis identifies a key frequency regime where the membrane-specific action of ethambutol leads to a change in the cellular dielectrophoretic response. This work represents the first such characterization of Mycobacteria and highlights the potential for DEP measurements to measure changes in mycobacterial membrane properties associated with chemical treatments or genetic mutation.
Asunto(s)
Electroforesis/métodos , Mycobacterium smegmatis/citología , Automatización , Impedancia Eléctrica , Modelos Biológicos , Estadística como AsuntoRESUMEN
We simulate electrothermally induced flow in polymeric, insulator-based dielectrophoresis (iDEP) systems with DC-offset, AC electric fields at finite thermal Péclet number, and we identify key regimes where electrothermal (ET) effects enhance particle deflection and trapping. We study a single, two-dimensional constriction in channel depth with parametric variations in electric field, channel geometry, fluid conductivity, particle electrophoretic (EP) mobility, and channel electroosmotic (EO) mobility. We report the effects of increasing particle EP mobility, channel EO mobility, and AC and DC field magnitudes on the mean constriction temperature and particle behavior. Specifically, we quantify particle deflection and trapping, referring to the deviation of particles from their pathlines due to dielectrophoresis as they pass a constriction and the stagnation of particles due to negative dielectrophoresis near a constriction, respectively. This work includes the coupling between fluid, heat, and electromagnetic phenomena via temperature-dependent physical parameters. Results indicate that the temperature distribution depends strongly on the fluid conductivity and electric field magnitude, and particle deflection and trapping depend strongly on the channel geometry. Electrothermal (ET) effects perturb the EO flow field, creating vorticity near the channel constriction and enhancing the deflection and trapping effects. ET effects alter particle deflection and trapping responses in insulator-based dielectrophoresis devices, especially at intermediate device aspect ratios (2 ≤ r ≤ 7) in solutions of higher conductivity (σ m ≥ 1 × 10(-3)S/m). The impact of ET effects on particle deflection and trapping are diminished when particle EP mobility or channel EO mobility is high. In almost all cases, ET effects enhance negative dielectrophoretic particle deflection and trapping phenomena.
Asunto(s)
Electroósmosis , Electroforesis , Modelos Químicos , Algoritmos , Fenómenos Químicos , Campos Electromagnéticos , Microfluídica , TermodinámicaRESUMEN
We demonstrate microfluidic devices for terahertz spectroscopy of biomolecules in aqueous solutions. The devices are fabricated out of a plastic material that is both mechanically rigid and optically transparent with near-zero dispersion in the terahertz frequency range. Using a lowpower terahertz time-domain spectrometer, we experimentally measure the absorption spectra of the vibrational modes of bovine serum albumin from 0.5 - 2.5 THz and find good agreement with previously reported data obtained using large-volume solutions and a high-power free-electron laser. Our results demonstrate the feasibility of performing high sensitivity terahertz spectroscopy of biomolecules in aqueous solutions with detectable molecular quantities as small as 10 picomoles using microfluidic devices.
Asunto(s)
Biopolímeros/análisis , Microfluídica/instrumentación , Espectrofotometría Infrarroja/instrumentación , Agua/química , Biopolímeros/química , Diseño de Equipo , Análisis de Falla de Equipo , Rayos Infrarrojos , Microfluídica/métodos , Microondas , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrofotometría Infrarroja/métodosRESUMEN
We present the development of a continuous-flow, "dielectrophoretic spectrometer" based on insulative DEP techniques and three-dimensional geometric design. Hot-embossed thermoplastic devices allow for high-throughput analysis and geometric control of electric fields via ridged microstructures patterned in a high width-to-depth aspect ratio (250:1) channel. We manipulate particles with dc-biased, ac electric fields and generate continuous-output streams of particles with a transverse outlet position specified by linear and nonlinear particle mobilities. We show, with simulation and experiment, that characteristic shape factors can be defined that capture the effects of constrictions in channel depth and that modulating the angle of these constrictions changes the resulting local DEP force. Microdevices are fabricated with an insulative constriction in channel depth, whose angle of incidence with the direction of flow varies continuously across the channel width. The resulting electric field gradients enable demonstration of a dielectrophoretic spectrometer that separates particles and controls their transverse channel position.