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Balsam J.,Office of Science and Engineering | Balsam J.,University of Maryland University College | Bruck H.A.,University of Maryland University College | Rasooly A.,Office of Science and Engineering | Rasooly A.,U.S. National Cancer Institute
Methods | Year: 2013

To overcome the limited sensitivity of phone cameras for mobile health (mHealth) fluorescent detection, we have previously developed a capillary array which enables a ~100× increase in detection sensitivity. However, for an effective detection platform, the optical configuration must allow for uniform measurement sensitivity between channels when using such a capillary array sensor. This is a challenge due to the parallax inherent in imaging long parallel capillary tubes with typical lens configurations.To enable effective detection, we have developed an orthographic projection. system in this work which forms parallel light projection images from the capillaries using an object-space telecentric lens configuration. This optical configuration results in a significantly higher degree of uniformity in measurement between channels, as well as a significantly reduced focal distance, which enables a more compact sensor.A plano-convex lens (f= 150. mm) was shown to produce a uniform orthographic projection. when properly combined with the phone camera's built in lens (f= 4. mm), enabling measurements of long capillaries (125. mm) to be made from a distance of 160. mm. The number of parallel measurements which can be made is determined by the size of the secondary lens. Based on these results, a more compact configuration with shorter 32. mm capillaries and a plano-convex lens with a shorter focal length (f= 10. mm) was constructed.This optical system was used to measure serial dilutions of fluorescein with a limit of detection (LOD) of 10. nM, similar to the LOD of a commercial plate reader. However, many plate readers based on standard 96 well plate requires sample volumes of 100. μl for measurement, while the capillary array requires a sample volume of less than 10. μl.This optical configuration allows for a device to make use of the ~100× increase in fluorescent detection sensitivity produced by capillary amplification while maintaining a compact size and capability to analyze extremely small sample volumes. Such a device based on a phone or other optical mHealth technology will have the sensitivity of a conventional plate reader but have greater mHealth clinical utility, especially for telemedicine and for resource-poor settings and global health applications. © 2013 . Source


Yang M.,University of Maryland Baltimore County | Yang M.,University of Jinan | Sun S.,University of Maryland Baltimore County | Kostov Y.,University of Maryland Baltimore County | And 2 more authors.
Sensors and Actuators, B: Chemical | Year: 2011

There is a well-recognized need for low cost biodetection technologies for resource-poor settings with minimal medical infrastructure. Lab-on-a-chip (LOC) technology has the ability to perform biological assays in such settings. The aim of this work is to develop a low cost, high-throughput detection system for the analysis of 96 samples simultaneously outside the laboratory setting. To achieve this aim, several biosensing elements were combined: a syringe operated ELISA lab-on-a-chip (ELISA-LOC) which integrates fluid delivery system into a miniature 96-well plate; a simplified non-enzymatic reporter and detection approach using a gold nanoparticle-antibody conjugate as a secondary antibody and silver enhancement of the visual signal; and carbon nanotubes (CNT) to increase primary antibody immobilization and improve assay sensitivity. Combined, these elements obviate the need for an ELISA washer, electrical power for operation and a sophisticated detector. We demonstrate the use of the device for detection of Staphylococcal enterotoxin B, a major foodborne toxin using three modes of detection, visual detection, CCD camera and document scanner. With visual detection or using a document scanner to measure the signal, the limit of detection (LOD) was 0.5 ng/ml. In addition to visual detection, for precise quantitation of signal using densitometry and a CCD camera, the LOD was 0.1 ng/ml for the CCD analysis and 0.5 ng/ml for the document scanner. The observed sensitivity is in the same range as laboratory-based ELISA testing. The point of care device can analyze 96 samples simultaneously, permitting high throughput diagnostics in the field and in resource poor areas without ready access to laboratory facilities or electricity. © 2011 Published by Elsevier B.V. Source


Bruck H.A.,University of Maryland College Park | Yang M.,Central South University | Kostov Y.,University of Maryland Baltimore County | Rasooly A.,Office of Science and Engineering | Rasooly A.,U.S. National Cancer Institute
Methods | Year: 2013

A new approach to label free biosensing has been developed based on the principle of "electrical percolation". In electrical percolation, long-range electrical connectivity is formed in randomly oriented and distributed systems of discrete elements. By applying this principle to biological interactions, it is possible to measure biological components both directly and electronically. The main element for electrical percolation biosensor is the biological semiconductor (BSC) which is a multi-layer 3-D carbon nanotube-antibody network. In the BSC, molecular interactions, such as binding of antigens to the antibodies, disrupt the network continuity causing increased resistance of the network. BSCs can be fabricated by immobilizing conducting elements, such as pre-functionalized single-walled carbon nanotubes (SWNTs)-antibody complex, directly onto a substrate, such as a Poly(methyl methacrylate) (PMMA) surface (also known as plexi-glass or Acrylic).BSCs have been demonstrated for direct (label-free) electronic measurements of antibody-antigen binding using SWNTs. If the concentration of the SWNT network is slightly above the electrical percolation threshold, then binding of a specific antigen to the pre-functionalized SWNT dramatically increases the electrical resistance due to changes in the tunneling between the SWNTs. Using anti-staphylococcal enterotoxin B (SEB) IgG as a "gate" and SEB as an "actuator", it was demonstrated that the BSC was able to detect SEB at concentrations of 1. ng/ml. Based on this concept, an automated configuration for BSCs is described here that enables real time continuous detection. The new BSC configuration may permit assembly of multiple sensors on the same chip to create "biological central processing units (CPUs)" with multiple biological elements, capable of processing and sorting out information on multiple analytes simultaneously. © 2013 . Source


Sun S.,Office of Science and Engineering | Sun S.,University of Maryland Baltimore County | Yang M.,University of Maryland Baltimore County | Yang M.,University of Jinan | And 3 more authors.
Lab on a Chip - Miniaturisation for Chemistry and Biology | Year: 2010

A miniature 96 sample ELISA-lab-on-a-chip (ELISA-LOC) was designed, fabricated, and tested for immunological detection of Staphylococcal Enterotoxin B (SEB). The chip integrates a simple microfluidics system into a miniature ninety-six sample plate, allowing the user to carry out an immunological assay without a laboratory. Assay reagents are delivered into the assay plate without the need for separate devices commonly used in immunoassays. The ELISA-LOC was constructed using Laminated Object Manufacturing (LOM) technology to assemble six layers with an acrylic (poly(methyl methacrylate) (PMMA)) core and five polycarbonate layers micromachined by a CO2 laser. The ELISA-LOC has three main functional elements: reagent loading fluidics, assay and detection wells, and reagent removal fluidics, a simple "surface tension" valve used to control the flow. To enhance assay sensitivity and to perform the assay without a lab, ELISA-LOC detection combines several biosensing elements: (1) carbon nanotube (CNT) technology to enhance primary antibody immobilization, (2) sensitive ECL (electrochemiluminescence) detection, and (3) a charge-coupled device (CCD) detector for measuring the light signal generated by ECL. Using a sandwich ELISA assay, the system detected SEB at concentrations as low as 0.1 ng ml-1, which is similar to the reported sensitivity of conventional ELISA. The fluidics system can be operated by a syringe and does not require power for operation. This simple point-of-care (POC) system is useful for carrying out various immunological assays and other complex medical assays without a laboratory. © 2010 The Royal Society of Chemistry. Source


Yang M.,University of Maryland, Baltimore | Yang M.,University of Jinan | Sun S.,University of Maryland, Baltimore | Kostov Y.,University of Maryland, Baltimore | And 2 more authors.
Lab on a Chip - Miniaturisation for Chemistry and Biology | Year: 2010

We describe a new eight channel Lab-On-a-Chip (LOC) for a Carbon Nanotube (CNT) based immunoassay with optical detection of Staphylococcal Enterotoxin B (SEB) for food safety applications. In this work, we combined four biosensing elements: (1) CNT technology for primary antibody immobilization, (2) Enhanced Chemiluminescence (ECL) for light signal generation, (3) a cooled charge-coupled device (CCD) for detection and (4) polymer lamination technology for developing a point of care immunological assay for SEB detection. Our concept for developing versatile LOCs, which can be used for many different applications, is to use a modular design with interchangeable recognition elements (e.g. various antibodies) to determine the specificity. Polymer lamination technology was used for the fabrication of a six layer, syringe operated LOC capable of analyzing eight samples simultaneously. An anti-SEB antibody-nanotube mixture was immobilized onto a polycarbonate strip, to serve as an interchangeable ligand surface that was then bonded onto the LOC. SEB samples are loaded into the device and detected by an ELISA assay using Horse Radish Peroxidase (HRP) conjugated anti-SEB IgG as a secondary antibody and ECL, with detection by a previously described portable cooled CCD detector. Eight samples of SEB in buffer or soy milk were assayed simultaneously with a limit of detection of 0.1 ng mL-1. CNT immobilization of the antibody increased the sensitivity of detection six fold. Use of a simple interchangeable immunological surface allows this LOC to be adapted to any immunoassay by simply replacing the ligand surface. A syringe was used to move fluids for this assay so no power is needed to operate the device. Our versatile portable point-of-care CCD detector combined with the LOC immunoassay method described here can be used to reduce the exposure of users to toxins and other biohazards when working outside the lab, as well as to simplify and increase sensitivity for many other types of immunological diagnostics and detection assays. © 2010 The Royal Society of Chemistry. Source

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