Droplet microfluidics for single-molecule and single-cell analysis in cancer research, diagnosis and therapy

Kang, Dong‐Ku; Ali, M. Monsur; Zhang, Kaixiang; Pone, Egest J.; Zhao, Weian

doi:10.1016/j.trac.2014.03.006

Cited by 105 publications

(63 citation statements)

References 55 publications

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“…Our rationale is that the confinement of samples in picoliter droplets can significantly increase the effective miRNA concentration and minimize the number of background molecules (interference) from plasma. 21 Therefore, microencapsulation permits high sensitivity and digital detection (i.e., each droplet has 0 or 1 target miRNA). This is supported by extensive previous droplet-based detection methods including ddPCR and digital RT-LAMP.…”

Section: Introductionmentioning

confidence: 99%

Digital quantification of miRNA directly in plasma using integrated comprehensive droplet digital detection

et al. 2015

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Quantification of miRNAs in blood can be potentially used for early disease detection, surveillance monitoring and drug response evaluation. However, quantitative and robust measurement of miRNAs in blood is still a major challenge in large part due to their low concentration and complicated sample preparation processes typically required in conventional assays. Here, we present the ‘Integrated Comprehensive Droplet Digital Detection’ (IC 3D) system where the plasma sample containing target miRNAs is encapsulated into microdroplets, enzymatically amplified and digitally counted using a novel, high-throughput 3D particle counter. Using Let-7a as a target, we demonstrate that IC 3D can specifically quantify target miRNA directly from blood plasma at extremely low concentrations ranging from 10s to 10,000 copies/mL in ≤ 3 hours without the need for sample processing such as RNA extraction. Using this new tool, we demonstrate that target miRNA content in colon cancer patient blood is significantly higher than that in healthy donor samples. Our IC 3D system has the potential to introduce a new paradigm for rapid, sensitive and specific detection of low-abundance biomarkers in biological samples with minimal sample processing.

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Section: Introductionmentioning

confidence: 99%

Digital quantification of miRNA directly in plasma using integrated comprehensive droplet digital detection

et al. 2015

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“…Accordingly, these compartments could proficiently provide a suitable home for CFPS reactions (see Figure 3). Additionally, artificial cells based on microbeads (Xiao et al, 2018), microgel (Jiao, Liu, Luo, Huck, & Yang, 2018), emulsions (Kang, Monsur Ali, Zhang, Pone, & Zhao, 2014), paper (Lim, Goh, & Khor, 2017), and vesicles (Elani, 2016) could be tailored for the generation of synthetic particles via microfluidic devices, which are further employed for the CFPS on the same chip (Damiati, Mhanna et al, 2018; Wang et al, 2018).…”

Section: Cfps: From Test Tube Reactions To Cell‐free Expression In MImentioning

confidence: 99%

Cell‐free protein synthesis: The transition from batch reactions to minimal cells and microfluidic devices

Ayoubi‐Joshaghani

Dianat‐Moghadam

Seidi

et al. 2020

Biotech & Bioengineering

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Thanks to the synthetic biology, the laborious and restrictive procedure for producing a target protein in living microorganisms by biotechnological approaches can now experience a robust, pliant yet efficient alternative. The new system combined with lab‐on‐chip microfluidic devices and nanotechnology offers a tremendous potential envisioning novel cell‐free formats such as DNA brushes, hydrogels, vesicular particles, droplets, as well as solid surfaces. Acting as robust microreactors/microcompartments/minimal cells, the new platforms can be tuned to perform various tasks in a parallel and integrated manner encompassing gene expression, protein synthesis, purification, detection, and finally enabling cell‐cell signaling to bring a collective cell behavior, such as directing differentiation process, characteristics of higher order entities, and beyond. In this review, we issue an update on recent cell‐free protein synthesis (CFPS) formats. Furthermore, the latest advances and applications of CFPS for synthetic biology and biotechnology are highlighted. In the end, contemporary challenges and future opportunities of CFPS systems are discussed.

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“…Cells trapped in droplets can be subjected to analysis or manipulation [32,33]. If the droplet contains enough nutrients, cells can be cultured within droplets [34] (Figure 2).…”

Section: Cell Biology and Tissue Engineeringmentioning

confidence: 99%

Droplet microfluidics: A tool for biology, chemistry and nanotechnology

Mashaghi

Abbaspourrad

Weitz

et al. 2016

TrAC Trends in Analytical Chemistry

311

188

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The ability to perform laboratory operations on small scales using miniaturized devices provides numerous benefits, including reduced quantities of reagents and waste as well as increased portability and controllability of assays. These operations can involve reaction components in the solution phase and as a result, their miniaturization can be accomplished through microfluidic approaches. One such approach, droplet microfluidics, provides a high-throughput platform for a wide range of assays and approaches in chemistry, biology and nanotechnology. We highlight recent advances in the application of droplet microfluidics in chipbased technologies, such as single-cell analysis tools, small-scale cell cultures, in-droplet chemical synthesis, high-throughput drug screening, and nanodevice fabrication.

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Droplet microfluidics for single-molecule and single-cell analysis in cancer research, diagnosis and therapy

Cited by 105 publications

References 55 publications

Digital quantification of miRNA directly in plasma using integrated comprehensive droplet digital detection

Digital quantification of miRNA directly in plasma using integrated comprehensive droplet digital detection

Cell‐free protein synthesis: The transition from batch reactions to minimal cells and microfluidic devices

Droplet microfluidics: A tool for biology, chemistry and nanotechnology

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