A microfluidic device for multiplexed protein detection in nano-liter volumes

Diercks, A.; Ozinsky, Adrian; Hansen, Carl L.; Spotts, James M.; Rodríguez, David; Aderem, Alan

doi:10.1016/j.ab.2008.12.012

Cited by 91 publications

(82 citation statements)

References 20 publications

(20 reference statements)

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“…Due to their miniaturization and automation, there are a number of advantages of using microfluidic systems, such as less sample/reagent consumption, reduced risk of contamination, less cost per analysis, lower power consumption, enhanced sensitivity and specificity, and higher reliability. Microfluidic systems have been developed for various biological analytical applications, such as DNA analysis [4][5][6][7][8], immunoassay [9][10][11][12][13], and cell analysis [14][15][16][17][18]. Moreover, a number of demonstrations showed that cell culture can be performed on the microfluidic systems to achieve higher throughput and more reliable results [19,20].…”

Section: Introductionmentioning

confidence: 99%

Review on Impedance Detection of Cellular Responses in Micro/Nano Environment

Lei

2014

Micromachines

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Abstract:In general, cell culture-based assays, investigations of cell number, viability, and metabolic activities during culture periods, are commonly performed to study the cellular responses under various culture conditions explored. Quantification of cell numbers can provide the information of cell proliferation. Cell viability study can understand the percentage of cell death under a specific tested substance. Monitoring of the metabolic activities is an important index for the study of cell physiology. Based on the development of microfluidic technology, microfluidic systems incorporated with impedance measurement technique, have been reported as a new analytical approach for cell culture-based assays. The aim of this article is to review recent developments on the impedance detection of cellular responses in micro/nano environment. These techniques provide an effective and efficient technique for cell culture-based assays.

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Section: Introductionmentioning

confidence: 99%

Review on Impedance Detection of Cellular Responses in Micro/Nano Environment

Lei

2014

Micromachines

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“…The reliable and high-sensitivity multiplexed protein detection is highly dependant on the quality of all the reagents, in particular, the antibodies. Compared with other methods for immunoassays using optically encoded microbeads [23,27], our method has several advantages. First, our encoding strategy is scalable and very easy to implement with readily available highquality microbeads.…”

Section: Multiplexed Protein Immunoassays and Sensitivitymentioning

confidence: 99%

“…If 10 encoded microbeads are used to interrogate a protein species of interest to provide 10-fold oversampling, 250 different protein species can be analysed within this 100 Â 100 mm area, the typical size of one spot used in conventional printed microarrays [16,45]. Therefore, our method will enable the integration of antibody array fabrication into microfluidic devices for sensitive multiplexed protein analysis [23,39,41,46] and potentially for single cell analysis.…”

Section: Multiplexed Protein Immunoassays and Sensitivitymentioning

confidence: 99%

“…The typical spots for each antibody type on a conventional microarray are relatively large (2500-10 000 mm 2 ). These spot sizes are not suitable for protein analysis of small sample volumes such as those from single cells [19,23]. Fortunately, these issues can be resolved by conjugating antibodies to encoded microbeads that are subsequently deposited onto a surface [24 -28].…”

Section: Introductionmentioning

confidence: 99%

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Multiplexed protein analysis using encoded antibody-conjugated microbeads

Theilacker

Roller

Barbee

et al. 2011

J. R. Soc. Interface.

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We describe a method for multiplexed analysis of proteins using fluorescently encoded microbeads. The sensitivity of our method is comparable to the sensitivity obtained by enzyme-linked immunosorbent assay while only 5 ml sample volumes are needed. Streptavidin-coated, 1 mm beads are encoded with a combination of fluorophores at different intensity levels. As a proof of concept, we demonstrate that 27 microbead populations can be readily encoded by affinity conjugation using three intensity levels for each of three different biotinylated fluorescent dyes. Four populations of encoded microbeads are further conjugated with biotinylated capture antibodies and then combined and immobilized in a microfluidic flow cell for multiplexed protein analysis. Using four uniquely encoded microbead populations, we show that a cancer biomarker and three cytokine proteins can be analysed quantitatively in the picogram per millilitre range by fluorescence microscopy in a single assay. Our method will allow for the fabrication of high density, bead-based antibody arrays for multiplexed protein analysis using integrated microfluidic devices and automated sample processing.

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“…Both samples and reagents can be added to the structures down to the nanolitre scale and transferred from one chamber to the next as they are going through a series of reactions. These steps can involve separation, mixing, incubation, washing steps and enzyme reactions with substrates necessary for the immunoassay [133]. It may also ease the automation of reactions alleviating the human error involved in the loss of accuracy and lowering the energy requirements of the assay [134].…”

Section: Miniaturisation Of Immunosensorsmentioning

confidence: 99%

Trends and Perspectives in Immunosensors

2012

Antibodies Applications and New Developments

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Immunosensors are devices that comprise both a biomolecular recognition system, such as an antibody and its corresponding antigen, and a transducer to translate the high affinity and specific binding event into a physical signal.Antibodies are produced by an immunological response to the presence of a foreign substance called an antigen. Antibodies may be immobilised onto a variety of platforms including bulk planar surfaces and nanoparticles by either covalent or adsorption strategies. Different interfaces between the bio-components and the detector are available to monitor in 'real-time' the signal generated by biological interactions. The transducers detect, for example, the change in electron transfer, absorbance, fluorescence, refractive index, mass change or heat transfer as the antibody binds to the antigen of interest. Such analytical devices have allowed a wide range of analytes to be identified and quantified such as pathogens, toxins, environmental food contaminants and disease biomarkers.The demand for sensitive, rapid, 'on-site' techniques has taken advantage of the latest advances in microfluidics and nanotechnology. This chapter will highlight current trends in immobilisation, micro/nano-fluidics/ and transducers utilised. A number of examples outlining the exploitation of these elements in immunosensors and their successful applications will be described.

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A microfluidic device for multiplexed protein detection in nano-liter volumes

Cited by 91 publications

References 20 publications

Review on Impedance Detection of Cellular Responses in Micro/Nano Environment

Review on Impedance Detection of Cellular Responses in Micro/Nano Environment

Multiplexed protein analysis using encoded antibody-conjugated microbeads

Trends and Perspectives in Immunosensors

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