RESUMEN
Quantifiable erectile dysfunction (ED) diagnosis involves the monitoring of rigidity and tumescence of the penile shaft during nocturnal penile tumescence (NPT). In this work, we introduce Erectile Dysfunction SENsor (EDSEN), a home-based wearable device for quantitative penile health monitoring based on stretchable microtubular sensing technology. Two types of sensors, the T- and R-sensors, are developed to effectively measure penile tumescence and rigidity, respectively. Conical models mimicking penile shaft were fabricated with polydimethylsiloxane (PDMS) material, using different base to curing agent ratios to replicate the different hardness properties of a penile shaft. A theoretical buckling force chart for the different penile models is generated to determine sufficiency criteria for sexual intercourse. An average erect penile length and circumference requires at least a Young's modulus of 179 kPa for optimal buckling force required for satisfactory sexual intercourse. The conical penile models were evaluated using EDSEN. Our results verified that the circumference of a penile shaft can be accurately measured by T-sensor and rigidity using the R-sensor. EDSEN provides a private and quantitative method to detect ED within the comfortable confines of the user's home.
Asunto(s)
Disfunción Eréctil , Dispositivos Electrónicos Vestibles , Masculino , Humanos , Disfunción Eréctil/diagnóstico , Erección Peniana , Dureza , Módulo de ElasticidadRESUMEN
Present work describes the methylene blue tagged thiolated aptamer-modified gold micro-array based biosensor for specific detection of IFN-γ. The microchips with the microelectrode array were fabricated using standard silicon microfabrication technologies, and modified with methylene blue tagged aptamer using standard gold thiol chemistry. Electrodes were characterized and tested using Cyclic Voltammetric (CV) and Square Wave Voltammetry (SQW) measurements in a standard three-electrode format at room temperature. On an aptamer modified electrode, aptamer density was estimated to be about 4.4 × 10(12)molecules/cm(2). In IFN-γ studies, oxidation peak currents were found to decrease and more than 50% signal suppression was achieved at 500 ng/ml. Further, the magnitude of signal suppression was found to be logarithmically proportional to the IFN-γ in the concentration range of 1-500 ng/ml, with a detection limit of 1.3 ng/ml (i.e. 0.8 fmol in used sample volume of 10 µl). Biosensor showed negligible signal changes (5%) in a very high non-specific protein background, while still able to differentiate target protein IFN-γ at 5 ng/ml. The results indicated that our sensor binds selectively to target molecules, and the non-specific binding where adsorption of BSA protein molecules may be effectively omitted from consideration.
Asunto(s)
Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Interferón gamma/aislamiento & purificación , Adsorción , Oro/química , Humanos , Interferón gamma/química , Límite de Detección , Azul de Metileno/químicaRESUMEN
In this report, we demonstrate a semi-integrated electrical biosensor for the detection of rare circulating tumor cells (CTCs) in blood. The sample was first enriched through a combination of immunomagnetic isolation and size filtration. The integration of both methods provided a high enrichment performance with a recovery rate above 70%, even for very low numbers of cancer cells present in the original sample (10 spiked MCF7 cells in 0.5 mL of blood). In the same system, the sample was then transferred to a microchip for further magnetic concentration, followed by immunochemical trapping and electronic detection by impedance spectroscopy. Three levels of spiked CTC number (30±2, 124±29, 273±23) in 10 µL of filtered blood sample were distinguished by monitoring the impedance change of the microelectrode array (MEA). The integration of different functions in a single system provided a methodology to process milliliter-sized blood samples at the macroscale and interface with the microdimensions of a highly sensitive electronic detector. The results showed that the whole system was able to detect different levels of spiked cancer cells without the use of time- and cost-intensive fluorescence labeling and image analysis. This has the potential to provide clinicians with a standalone system to monitor changes in CTC numbers throughout therapy conveniently and frequently for efficient cancer treatments.
Asunto(s)
Técnicas Biosensibles/instrumentación , Neoplasias de la Mama/sangre , Neoplasias de la Mama/fisiopatología , Conductometría/instrumentación , Células Neoplásicas Circulantes , Línea Celular Tumoral , Diseño de Equipo , Análisis de Falla de Equipo , HumanosRESUMEN
Lateral patch-clamping has emerged as a chip-based platform for automation of the conventional patch-clamp technique, the 'gold' standard for studying cellular ion channels. The conventional technique, as it relies on skilled-maneuver of glass micropipettes to patch cells, is extremely delicate, low in throughput, and thus cannot be used for primary screening of compounds against ion channels. Direct integration of glass capillaries on silicon provides lateral junctions for automated trapping and patching of cells. We demonstrate here a method of scaling up the lateral junctions to a standard 1536-well microtiter plate format. A single unit of 1536-well plate has been formed here on a 9 mm by 9 mm microstructured silicon with the inclusive of 16 wells molded in a capping layer made of polydimethylsiloxane (PDMS). The silicon substrate provides integrated glass capillaries (total 12) and their associated microfluidic network. Each glass capillary has an independent access through a dedicated well in PDMS and leads to a centralized channel in which cell suspension can be delivered through one of the remaining 4 wells. The unit has been tested on RBL-1 cells by recording whole-cell activity from inwardly rectifying endogenous potassium channels. A revised test protocol has been prescribed to avoid inaccurate readings due to altered ionic composition of the recording buffer when a typical suction is applied to capture cells.
Asunto(s)
Potenciales de Acción/fisiología , Técnicas de Cultivo de Célula/instrumentación , Electrodos , Potenciales de la Membrana/fisiología , Técnicas Analíticas Microfluídicas/instrumentación , Neuronas/fisiología , Técnicas de Placa-Clamp/instrumentación , Línea Celular , Diseño de Equipo , Análisis de Falla de Equipo , Humanos , Técnicas de Placa-Clamp/métodosRESUMEN
This paper presents a novel microfluidic system for rapid label-free detection of endothelial progenitor cells (EPCs) from small volumes of white blood cells samples, to obtain a bedside cardiovascular diagnostic solution. The system was built on a single 1 cm(2) microelectrode array silicon chip, integrated with negative dielectrophoresis for cell trapping, surface immunochemistry for selective cell capture, and fluidics for cell washing and impedance detection. The level of circulating EPC level in blood is a biomarker of clinical interest, linked to the assessment of risk factors in cardiovascular diseases which are a major global concern. Rare EPCs are usually detected through in vitro culture or flow cytometry, which are too time-consuming to bring timely reports in acute diseases. Although microfluidics approaches have enabled reduced processing time and enhanced portability, their sensitivity and processing volumes are still inadequate for rare cell detection at a bedside setting. Using small highly sensitive microelectrodes, our novel integrated system achieved the detection of 720 EPCs in a small 12 microl sample of 72,000 peripheral blood mononuclear cells (PBMC), i.e. equivalent to a concentration of EPCs of 0.1% of 100 microl blood. This demonstrated that clinically significant level of EPCs (<0.5% of PBMC) could be detected for the first time on a detection system at bedside set-up, showing great potential in applications for point-of-care diagnosis.
Asunto(s)
Recuento de Células/instrumentación , Separación Celular/instrumentación , Células Endoteliales/citología , Inmunoensayo/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Sistemas de Atención de Punto , Células Madre/citología , Técnicas Biosensibles/instrumentación , Electroforesis/instrumentación , Diseño de Equipo , Análisis de Falla de Equipo , Humanos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Coloración y EtiquetadoRESUMEN
We describe a self-contained fully-enclosed cartridge for lab-on-a-chip applications where sample and reagents can be applied sequentially as is performed in a heterogeneous immunoassay, or nucleic acid extraction. Both the self-contained and fully-enclosed features of the cartridge are sought to ensure its safe use in the field by unskilled staff. Simplicity in cartridge design and operation is obtained via adopting a valveless concept whereby reagents are stored and used in the form of liquid plugs isolated by air spacers around a fluidic loop. Functional components integrated in the loop include a microfluidic chip specific to the target application, a novel peristaltic pump to displace the liquid plugs, and a pair of removable tubing segments where one is used to introduce biological sample and while the other is to collect eluant. The novel pump is fabricated through soft-lithography technique and works by pinching a planar channel under stainless-steel ball bearings that have been magnetically loaded. The utility of the cartridge is demonstrated for automated extraction and purification of nucleic acids (DNA) from a cell lysate on a battery-operated portable system. The cartridge shown here can be further extended to sample-in-answer-out diagnostic tests.
Asunto(s)
Técnicas Analíticas Microfluídicas/instrumentación , ADN/aislamiento & purificación , Reacción en Cadena de la PolimerasaRESUMEN
We demonstrate a simple planar peristaltic pump fabricated in poly(dimethylsiloxane) (PDMS) via soft lithography and suitable for microfluidic integration.
Asunto(s)
Equipos Desechables , Bombas de Infusión , Técnicas Analíticas Microfluídicas/instrumentación , Dimetilpolisiloxanos/química , Técnicas Analíticas Microfluídicas/métodos , Siliconas/químicaRESUMEN
High-throughput screening of drug candidates for channelopathies can greatly benefit from an automated patch-clamping assay. Automation of the patch clamping through microfluidics ideally requires on-chip integration of glass capillaries with substantially round cross section. Such round capillaries, if they can only be integrated to connect isolated reservoirs on a substrate surface, will lead to a "lateral" configuration which is simple yet powerful for the patch clamping. We demonstrate here "lateral" patch clamping through microfluidic integration of substantially round glass capillaries in a novel process. The process adopts two well-known phenomena from microelectronics: keyhole-void formation and thermal-reflow of phosphosilicate glass in silicon trenches. The process relies on the same physical principle as the preparation of conventional micropipette electrodes by heat-pulling and fire-polishing glass tubes. The optimized process forms capillaries with a diameter approximately 1.5 microm and variation <10%. Functionality of the integrated glass capillaries for the patch-clamp recording has been verified by statistical test results from a sample of one hundred capillaries on mammalian cells (RBL-1) in suspension: 61% formed gigaseals (>1 GOmega) and of those approximately 48% (29% of all) achieved whole-cell recordings. Pharmacological blockade of ion channel activity and longevity of a whole-cell mode on these capillaries have also been presented.