Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms

Zhang, Qiang; Wang, Tingting; Zhou, Qian; Zhang, Peng; Gong, Yanhai; Gou, Honglei; Xu, Jian; Ma, Bo

doi:10.1038/srep41192

Cited by 72 publications

(83 citation statements)

References 46 publications

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“…To increase the operation resolution to single‐droplet scale, a facile microfluidic platform was introduced to collect individual droplets . With a microscope to identify the arrival of positive droplets and a movable stage to track its movement, single droplet is manually sorted into dispensing channel through solenoid valve suction.…”

Section: Introductionmentioning

confidence: 99%

On‐Demand Droplet Collection for Capturing Single Cells

Nan

Lai

Tang

et al. 2019

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Droplet‐based microfluidic techniques are extensively used in efficient manipulation and genome‐wide analysis of individual cells, probing the heterogeneity among populations of individuals. However, the extraction and isolation of single cells from individual droplets remains difficult due to the inevitable sample loss during processing. Herein, an automated system for accurate collection of defined numbers of droplets containing single cells is presented. Based on alternate sorting and dispensing in three branch channels, the droplet number can be precisely controlled down to single‐droplet resolution. While encapsulating single cells and reserving one branch as a waste channel, sorting can be seamlessly integrated to enable on‐demand collection of single cells. Combined with a lossless recovery strategy, this technique achieves capture and culture of individual cells with a harvest rate of over 95%. The on‐demand droplet collection technique has great potential to realize quantitative processing and analysis of single cells for elucidating the role of cell‐to‐cell variations.

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Section: Introductionmentioning

confidence: 99%

On‐Demand Droplet Collection for Capturing Single Cells

Nan

Lai

Tang

et al. 2019

Small

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“…(d) (i) On‐chip monodispersed droplets for single‐cell isolation, reprinted with permission from Ref. . Copyright 2017 Springer Nature, and (ii) drop on‐demand inkjet printing of single‐cell in droplet, reprinted with permission from Ref.…”

Section: Single‐cell Isolationmentioning

confidence: 99%

“…The mSCP can isolate single cells from 5–10 cells per μL of cell suspension droplet and compatible with the microscopic identification of isolated single‐cells. Recently, Zhang and co‐workers developed a droplet microfluidic device by integrating cell encapsulation, droplet inspection, single‐cell droplet sorting and exporting on one chip (Figure di). Chen et al .…”

Section: Single‐cell Isolationmentioning

confidence: 99%

Microfluidic Devices in the Fast‐Growing Domain of Single‐Cell Analysis

Khan

Mao

et al. 2018

Chemistry A European J

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Recent advances in cellular analysis revealed that the seemingly identical cells are heterogeneous in term of functionality, compositions, and genetic performance. These differences cause difficulty in the diagnostic for a specific model of disease. Detection of biomolecules such as DNA, RNA, and protein or analysis of cell(s), detection of cell surface molecules, and secreted protein, can help us to improve the understanding of a targeted disease and development of new diagnostic and therapeutic approaches. A single-cell includes the minute quantity of these target molecules. Microfluidic devices have the ability to capture a single-cell and its lysate into a pico or femtoliter volumes droplet, or micro-well thus preventing dilution and limiting diffusion. In this review we described the advancement and limitations in microfluidic techniques used toward single-cells analysis.

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“…A commercial microfluidic circuitry chip DEPArray System (Menarini Silicon Biosystems, Inc.) containing an array of individually controllable electrodes to create a dielectrophoretic (DEP) cage around each cell for single CTC isolation is also available [65] . Besides isolation of CTCs from blood, the microfluidic platform can also be used for single-cell isolation from other tissues [66,67] . For example, an innovative workflow using DEPArray system was established to examine tumor heterogeneity using FFPE samples, providing a solution for genetic analysis using minute archival clinical samples [68] .…”

Section: Microfluidic-based Cell Isolationmentioning

confidence: 99%

Unmasking tumor heterogeneity and clonal evolution by single-cell analysis

Shi¹,

Chakraborty²,

Chaudhuri³

2018

JCMT

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The intratumoral heterogeneity orchestrated by the tumor intrinsic and extrinsic mechanisms enable cancers to persist and spread notwithstanding the use of aggressive interventional therapies. The heterogeneity is revealed at multiple levels-at the level of individual tumor cells, in the cellular composition of tumor infiltrates and in the chemical microenvironment in which the cells reside. Deconvoluting the complex nature of the cell types present in the tumor, along with the homo and heterotypic interactions between different cell types can produce novel insights of biological and clinical relevance. However, most techniques analyze tumors at a gross level missing key inter-cell-type genotypic and phenotypic differences. The advent of single-cell sequencing has given an unprecedented opportunity to analyze the tumor at a resolution that not only captures the diversity of the cellular composition of a tumor but also provides information on the genetic, epigenetic and functional states of different cell types. In this review, we summarize the genesis of tumor heterogeneity, its impact on tumor growth and progression and their clinical consequences. We present an overview of the currently available platforms for isolation and sequencing of single tumor cells and provide evidence of its utility in precision medicine and personalized therapy.

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Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms

Cited by 72 publications

References 46 publications

On‐Demand Droplet Collection for Capturing Single Cells

On‐Demand Droplet Collection for Capturing Single Cells

Microfluidic Devices in the Fast‐Growing Domain of Single‐Cell Analysis

Unmasking tumor heterogeneity and clonal evolution by single-cell analysis

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