RESUMO
A dielectrophoretic device employing a planar array of microelectrodes is designed for controlled transport of individual microparticles. By exciting the electrodes in sequence, a moving dielectrophoretic force is created that can drag a particle across the electrodes in a straight line. The electrode shapes are designed to counter any lateral drift of the trapped particle during transport. This facilitates single particle transport by creating a narrow two-dimensional corridor for the moving dielectrophoretic force to operate on. The design and analysis processes are discussed in detail. Numerical simulations are performed to calculate the electromagnetic field distribution and the generated dielectrophoretic force near the electrodes. The Langevin equation is used for analyzing the trajectory of a microparticle under the influence of the external forces. The simulations show how the designed electrode geometry produces the necessary lateral confinement required for successful particle transport. Finally, experimental results are presented showing controlled bidirectional linear transport of single polystyrene beads of radius 10 and 5 µm for a distances 840 and 1100 µm, respectively. The capabilities of the proposed platform make it suitable for micro total analysis systems (µTAS) and lab-on-a-chip (LOC) applications.
Assuntos
Dispositivos Lab-On-A-Chip , Poliestirenos , Microeletrodos , Eletroforese/métodosRESUMO
To achieve the mission of personalized medicine, centering on delivering the right drug to the right patient at the right dose, therapeutic drug monitoring solutions are necessary. In that regard, wearable biosensing technologies, capable of tracking drug pharmacokinetics in noninvasively retrievable biofluids (e.g., sweat), play a critical role, because they can be deployed at a large scale to monitor the individuals' drug transcourse profiles (semi)continuously and longitudinally. To this end, voltammetry-based sensing modalities are suitable, as in principle they can detect and quantify electroactive drugs on the basis of the target's redox signature. However, the target's redox signature in complex biofluid matrices can be confounded by the immediate biofouling effects and distorted/buried by the interfering voltammetric responses of endogenous electroactive species. Here, we devise a wearable voltammetric sensor development strategy-centering on engineering the molecule-surface interactions-to simultaneously mitigate biofouling and create an "undistorted potential window" within which the target drug's voltammetric response is dominant and interference is eliminated. To inform its clinical utility, our strategy was adopted to track the temporal profile of circulating acetaminophen (a widely used analgesic and antipyretic) in saliva and sweat, using a surface-modified boron-doped diamond sensing interface (cross-validated with laboratory-based assays, R2 â¼ 0.94). Through integration of the engineered sensing interface within a custom-developed smartwatch, and augmentation with a dedicated analytical framework (for redox peak extraction), we realized a wearable solution to seamlessly render drug readouts with minute-level temporal resolution. Leveraging this solution, we demonstrated the pharmacokinetic correlation and significance of sweat readings.
Assuntos
Acetaminofen/análise , Monitoramento de Medicamentos/métodos , Saliva/química , Suor/química , Acetaminofen/administração & dosagem , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Monitoramento de Medicamentos/instrumentação , Humanos , Medicina de Precisão , Dispositivos Eletrônicos VestíveisRESUMO
Optimization of targeted cell capture with microfluidic devices continues to be a challenge. On the one hand, microfluidics allow working with microliter volumes of liquids, whereas various applications in the real world require detection of target analyte in large volumes, such as capture of rare cell types in several ml of blood. This contrast of volumes (microliter vs. ml) has prevented the emergence of microfluidic cell capture sensors in the clinical setting. Here, we study the improvement in cell capture and throughput achieved using parallel bioactivated microfluidic channels. The device consists of channels in parallel with each other tied to a single channel. We discuss fabrication and testing of our devices, and show the ability for an improvement in throughput detection of target cells.
Assuntos
Separação Celular/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Animais , Anticorpos Imobilizados/química , Anticorpos Imobilizados/metabolismo , Candida albicans/citologia , Dimetilpolisiloxanos/química , Saccharomyces cerevisiae/citologia , Fatores de TempoRESUMO
The present study reports on the retention of conformational flexibility of a model allosteric protein upon immobilization on self-assembled monolayers (SAMs) on gold. Organothiolated SAMs of different compositions were utilized for adsorptive and covalent attachment of bovine liver glutamate dehydrogenase (GDH), a well-characterized allosteric enzyme. Sensitive fluorimetric assays were developed to determine immobilization capacity, specific activity, and allosteric properties of the immobilized preparations as well as the potential for repeated use and continuous catalytic transformations. The allosteric response of the free and immobilized forms towards ADP, L-leucine and high concentrations of NAD(+), some of the well-known activators for this enzyme, were determined and compared. The enzyme immobilized by adsorption or chemical binding responded similarly to the activators with a greater degree of activation, as compared to the free form. Also loss of activity involving the two immobilization procedures were similar, suggesting that residues essential for catalytic activity or allosteric properties of GDH remained unchanged in the course of chemical modification. A recently established method was used to predict GDH orientation upon immobilization, which was found to explain some of the experimental results presented. The general significance of these observations in connection with retention of native properties of protein structures upon immobilization on SAMs is discussed.
Assuntos
Materiais Revestidos Biocompatíveis/química , Glutamato Desidrogenase/química , Glutamato Desidrogenase/ultraestrutura , Ouro/química , Modelos Químicos , Modelos Moleculares , Adsorção , Animais , Sítios de Ligação , Bovinos , Simulação por Computador , Ativação Enzimática , Estabilidade Enzimática , Enzimas Imobilizadas/química , Ligação Proteica , Conformação ProteicaRESUMO
This paper presents the modeling of the electrical properties of bioactivated nanopores, customized nanopore devices with a biological macromolecule attached in the pore as the probe. These devices are capable of detecting and analyzing interactions between the attached biomolecule and the molecules in the analyte at a single molecule level.
Assuntos
Biopolímeros/química , Técnicas Biossensoriais/instrumentação , Membranas Artificiais , Modelos Químicos , Técnicas de Sonda Molecular/instrumentação , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Bioensaio/instrumentação , Bioensaio/métodos , Biopolímeros/análise , Técnicas Biossensoriais/métodos , Simulação por Computador , Desenho Assistido por Computador , Eletroquímica/instrumentação , Eletroquímica/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Microeletrodos , PorosidadeRESUMO
This paper presents a novel device which provides the opportunity to perform high-throughput biochemical assays on different individual cells. In particular, the proposed device is suited to screen the rare cells in biological samples for early stage cancer diagnosis and explore their biochemical functionality. In the process, single cells are precisely positioned and captured in activated micropores. To show the performance of the proposed device, cultured yeast cells and human epithelial circulating tumor cells are successfully captured.