Multiplexed microfluidic blotting of proteins and nucleic acids by parallel, serpentine microchannels

He, Sha; Zhang, Yi; Wang, Pei; Xu, Xingzhi; Zhu, Kui; Pan, Wenying; Liu, Wenwen; Cai, Kaiyong; Sun, Jiashu; Zhang, Wei; Jiang, Xingyu

doi:10.1039/c4lc00901k

Cited by 22 publications

(19 citation statements)

References 33 publications

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“…In this pursuit, Rane et al developed a high-throughput, continuous-flow droplet platform for detection of bacterial DNA (Rane, TD; Chen, L; Zec, HC; Wang, 2014). Here, loop-mediated isothermal amplification (LAMP) was used for amplifying and detecting single copies of target genes purified from samples containing Neisseria gonorrhea (Figure 4C).…”

Section: Detection and Screening Of Nucleic Acid Biomarkersmentioning

confidence: 99%

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Droplet microfluidics for high‐sensitivity and high‐throughput detection and screening of disease biomarkers

Kaushik

Hsieh

Wang

2018

WIREs Nanomed Nanobiotechnol

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Biomarkers are nucleic acids, proteins, single cells, or small molecules in human tissues or biological fluids whose reliable detection can be used to confirm or predict disease and disease states. Sensitive detection of biomarkers is therefore critical in a variety of applications including disease diagnostics, therapeutics, and drug screening. Unfortunately for many diseases, low abundance of biomarkers in human samples and low sample volumes render standard benchtop platforms like 96-well plates ineffective for reliable detection and screening. Discretization of bulk samples into a large number of small volumes (fL-nL) via droplet microfluidic technology offers a promising solution for high-sensitivity and high-throughput detection and screening of biomarkers. Several microfluidic strategies exist for high-throughput biomarker digitization into droplets, and these strategies have been utilized by numerous droplet platforms for nucleic acid, protein, and single-cell detection and screening. While the potential of droplet-based platforms has led to burgeoning interest in droplets, seamless integration of sample preparation technologies and automation of platforms from biological sample to answer remain critical components that can render these platforms useful in the clinical setting in the near future. This article is categorized under: Diagnostic Tools > Biosensing Diagnostic Tools > Diagnostic Nanodevices Therapeutic Approaches and Drug Discovery > Emerging Technologies Therapeutic Approaches and Drug Discovery > Nanomedicine for Infectious Disease.

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Section: Detection and Screening Of Nucleic Acid Biomarkersmentioning

confidence: 99%

“…Critically, the assay was conducted in continuous-flow in an integrated platform that facilitated digitization of targets, on-chip incubation, and detection. (Reproduced from (Rane, TD; Chen, L; Zec, HC; Wang, 2014) with permission of The Royal Society of Chemistry).…”

Section: Figurementioning

confidence: 99%

Droplet microfluidics for high‐sensitivity and high‐throughput detection and screening of disease biomarkers

Kaushik

Hsieh

Wang

2018

WIREs Nanomed Nanobiotechnol

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“…28 As shown in Fig. 1, the PDMS chip incorporated seven parallel channels designed to weave along multiple lanes in a PVDF membrane, sharing one set of inlet and outlet ports.…”

Section: Multiplexed Microfluidic Probing Following Western Blottingmentioning

confidence: 99%

Recent advances in microscale western blotting

2016

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Western blotting is a ubiquitous tool used extensively in the clinical and research settings to identify proteins and characterize their levels. It has rapidly become a mainstay in research laboratories due to its specificity, low cost, and ease of use. The specificity arises from the orthogonal processes used to identify proteins. Samples are first separated based on size and then probed with antibodies specific for the protein of interest. This confirmatory approach helps avoid pitfalls associated with antibody cross-reactivity and specificity issues. While the technique has evolved since its inception, the last decade has witnessed a paradigm shift in Western blotting technology. The introduction of capillary and microfluidic platforms has significantly decreased time and sample requirements while enabling high-throughput capabilities. These advances have enabled Western analysis down to the single cell level in highly parallel formats, opening vast new opportunities for studying cellular heterogeneity. Recent innovations in microscale Western blotting are surveyed, and the potential for enhancing detection using advances in label-free biosensing is briefly discussed.

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“…[19] To realize multiplexed blotting, a parallel, serpentine microfluidic channels chip can dramatically improve the WB system (Figure 1f). [20] This multiplexed micro-blotting Figure 1. PDMS-based microfluidics immunoassay chips.…”

Section: Reviewmentioning

confidence: 99%

“…Reproduced with permission. [20] Microfluidic immunoassay can detect multiplex biomarkers simultaneous. However the low efficiency in the downstream data analysis still waste a lot of time.…”

Section: Reviewmentioning

confidence: 99%

Materials for Microfluidic Immunoassays: A Review

Mou

Jiang

2017

Adv Healthcare Materials

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117

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Conventional immunoassays suffer from at least one of these following limitations: long processing time, high costs, poor user‐friendliness, technical complexity, poor sensitivity and specificity. Microfluidics, a technology characterized by the engineered manipulation of fluids in channels with characteristic lengthscale of tens of micrometers, has shown considerable promise for improving immunoassays that could overcome these limitations in medical diagnostics and biology research. The combination of microfluidics and immunoassay can detect biomarkers with faster assay time, reduced volumes of reagents, lower power requirements, and higher levels of integration and automation compared to traditional approaches. This review focuses on the materials‐related aspects of the recent advances in microfluidics‐based immunoassays for point‐of‐care (POC) diagnostics of biomarkers. We compare the materials for microfluidic chips fabrication in five aspects: fabrication, integration, function, modification and cost, and describe their advantages and drawbacks. In addition, we review materials for modifying antibodies to improve the performance of the reaction of immunoassay. We also review the state of the art in microfluidic immunoassays POC platforms, from the laboratory to routine clinical practice, and also commercial products in the market. Finally, we discuss the current challenges and future developments in microfluidic immunoassays.

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Multiplexed microfluidic blotting of proteins and nucleic acids by parallel, serpentine microchannels

Cited by 22 publications

References 33 publications

Droplet microfluidics for high‐sensitivity and high‐throughput detection and screening of disease biomarkers

Droplet microfluidics for high‐sensitivity and high‐throughput detection and screening of disease biomarkers

Recent advances in microscale western blotting

Materials for Microfluidic Immunoassays: A Review

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