A Self‐Digitization Dielectrophoretic (SD‐DEP) Chip for High‐Efficiency Single‐Cell Capture, On‐Demand Compartmentalization, and Downstream Nucleic Acid Analysis

Qin, Yuling; Wu, Li; Schneider, Thomas; Yen, Gloria S.; Wang, Jiasi; Xu, Shihan; Li, Min; Paguirigan, Amy L.; Smith, Jordan L.; Radich, Jerald P.; Anand, Robbyn K.; Chiu, Daniel T.

doi:10.1002/ange.201807314

Cited by 13 publications

(7 citation statements)

References 44 publications

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“…Loop-mediated isothermal amplification (LAMP) is an ultrasensitive gene amplification technique that is carried out at a constant temperature without the requirement of a thermal cycler. − Compared with other isothermal amplification techniques, such as nucleic acid sequence-based amplification (NASBA), strand displacement amplification (SDA), recombinase polymerase amplification (RPA), and helicase-dependent amplification (HDA), LAMP can achieve 10 9 -fold amplification efficiency in less than 1 h using a single polymerase at a constant temperature. ,, In addition, LAMP also has advantages of other amplification methods such as simplicity, time saving, and low cost. − …”

mentioning

confidence: 99%

CUT-LAMP: Contamination-Free Loop-Mediated Isothermal Amplification Based on the CRISPR/Cas9 Cleavage

et al. 2020

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Loop-mediated isothermal amplification (LAMP) is a sensitive and widely used gene amplification technique. However, high amplification efficiency and amplification products containing multiple inverted repeats make the LAMP reaction extremely vulnerable to false-positive amplification caused by contamination. Herein, a contamination-free LAMP (CUT-LAMP) assisted by the CRISPR/Cas9 cleavage with superior reliability and durability has been reported. The core of CUT-LAMP is the engineering of the forward or backward inner primer in the target-independent region, which makes the LAMP products contain a protospacer adjacent motif (PAM) site for the CRISPR/Cas9 recognition. For the CUT-LAMP reaction, cross-contamination can be efficiently cleaved by the corresponding Cas9/sgRNA, but the target gene can get rid of digestion due to the lack of a PAM site near the recognition region. CUT-LAMP shows impressive contamination resistance but does not significantly increase procedure complexity; thus, it represents a simple and versatile toolkit facilitating the adoption by open-and closed-tube detection format.

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mentioning

confidence: 99%

CUT-LAMP: Contamination-Free Loop-Mediated Isothermal Amplification Based on the CRISPR/Cas9 Cleavage

et al. 2020

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“…In addition, DEP-based methods have recently demonstrated great capacity for single-CTC manipulation. As Figure b displayed, Qin and co-workers developed a self-digitization dielectrophoretic (SD-DEP) chip for single-CTC manipulation, together with downstream nucleic acid analysis . Their data showed that single CTCs were captured, immobilized, lysed, and analyzed within one portable and integrated device.…”

Section: Microfluidic Technologies For Ctc Isolationmentioning

confidence: 99%

Section: Microfluidic Technologies For Ctc Isolationmentioning

confidence: 99%

Emerging Microfluidic Technologies for the Detection of Circulating Tumor Cells and Fetal Nucleated Red Blood Cells

Wei

Chen

Guo

et al. 2021

ACS Appl. Bio Mater.

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Blood tests have been a powerful tool for the clinical analysis of many diseases. With the advances in microfluidic technology, two more specific indicators from the circulation system, namely, emerging "liquid biopsy" of circulating tumor cells (CTCs) and fetal nucleated red blood cells (fNRBCs), can be screened and analyzed as a simple blood test for the noninvasive diagnosis of cancers as well as fetal disorders. The unique feature of precisely manipulating a trace of fluid endows microfluidic devices with the ability to isolate CTCs or fNRBCs from numerous blood cells with high performance, which undoubtedly facilitates biomedical applications of these two kinds of rare cells. In this review, advanced developments in microfluidic technologies focusing on the detection and sorting of rare CTCs and fNRBCs from peripheral blood are summarized. The development of microfluidic devices incorporated with various multifunctional microstructures and nanomaterials for enhancing the sensitivity, purity, and viability of CTC or fNRBC detection enables CTC molecular analysis and fNRBC-based noninvasive prenatal diagnosis (NIPD). These microfluidics-based approaches provide great potential opportunities in noninvasive cancer diagnosis or NIPD applications.

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“…Silicon masters were created by using standard photolithographic techniques described previously. 43 SU-8 2050 photoresist (MicroChem, Westborough, MA) was used for spin coating. To fabricate microfluidic chips, polydimethylsiloxane (PDMS) with a 1:10 ratio of precursor to polymer base was cured and sealed to a glass substrate immediately after exposure to O 2 plasma for 30 s. If not used immediately, chips were covered and stored for up to one month until use.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…Silicon masters were created by using standard photolithographic techniques described previously . SU-8 2050 photoresist (MicroChem, Westborough, MA) was used for spin coating.…”

Section: Experimental Sectionmentioning

confidence: 99%

Sequential Ensemble-Decision Aliquot Ranking Isolation and Fluorescence In Situ Hybridization Identification of Rare Cells from Blood by Using Concentrated Peripheral Blood Mononuclear Cells

Qin

et al. 2021

Anal. Chem.

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Isolation and analysis of circulating rare cells is a promising approach for early detection of cancer and other diseases and for prenatal diagnosis. Isolation of rare cells is usually difficult due to their heterogeneity as well as their low abundance in peripheral blood. We previously reported a two-stage ensemble-decision aliquot ranking platform (S-eDAR) for isolating circulating tumor cells from whole blood with high throughput, high recovery rate (>90%), and good purity (>70%), allowing detection of low surface antigen-expressing cancer cells linked to metastasis. However, due to the scarcity of these cells, large sample volumes and large quantities of antibodies were required to isolate sufficient cells for downstream analysis. Here, we drastically increased the number of nucleated cells analyzed by first concentrating peripheral blood mononuclear cells (PBMCs) from whole blood by density gradient centrifugation. The S-eDAR platform was capable of isolating rare cells from concentrated PBMCs (108/mL, equivalent to processing ∼20 mL of whole blood in the 1 mL sample volume used by our instrument) at a high recovery rate (>85%). We then applied the S-eDAR platform for isolating rare fetal nucleated red blood cells (fNRBCs) from concentrated PBMCs spiked with umbilical cord blood cells and confirmed fNRBC recovery by immunostaining and fluorescence in situ hybridization, demonstrating the potential of the S-eDAR system for isolating rare fetal cells from maternal PBMCs to improve noninvasive prenatal diagnosis.

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A Self‐Digitization Dielectrophoretic (SD‐DEP) Chip for High‐Efficiency Single‐Cell Capture, On‐Demand Compartmentalization, and Downstream Nucleic Acid Analysis

Cited by 13 publications

References 44 publications

CUT-LAMP: Contamination-Free Loop-Mediated Isothermal Amplification Based on the CRISPR/Cas9 Cleavage

CUT-LAMP: Contamination-Free Loop-Mediated Isothermal Amplification Based on the CRISPR/Cas9 Cleavage

Emerging Microfluidic Technologies for the Detection of Circulating Tumor Cells and Fetal Nucleated Red Blood Cells

Sequential Ensemble-Decision Aliquot Ranking Isolation and Fluorescence In Situ Hybridization Identification of Rare Cells from Blood by Using Concentrated Peripheral Blood Mononuclear Cells

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