Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis

Wen, Na; Zhao, Zhan; Fan, Beiyuan; Chen, Deyong; Men, Dong; Wang, Junbo; Chen, Jian

doi:10.3390/molecules21070881

Cited by 90 publications

(53 citation statements)

References 93 publications

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“…Microfluidic sorting of hydrogel microcapsules enables segregation of subpopulations of microcapsules of particular interest, purification of heterogeneously mixed microcapsules, and manipulation of individual microcapsules 126, 127 . Sorting techniques are classified as either “active” or “passive”.…”

Section: On-chip Manipulations For Cell-laden Hydrogel Microcapsulesmentioning

confidence: 99%

Generation and manipulation of hydrogel microcapsules by droplet-based microfluidics for mammalian cell culture

et al. 2017

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Hydrogel microcapsules provide miniaturized and biocompatible niches for three-dimensional (3D) in vitro cell culture. They can be easily generated by droplet-based microfluidics with tunable size, morphology, and biochemical properties. Therefore, microfluidic generation and manipulation of cell-laden microcapsules can be used for 3D cell culture to mimic the in vivo environment towards applications in tissue engineering and high throughput drug screening. In this review of recent advances mainly since 2010, we will first introduce general characteristics of droplet-based microfluidic devices for cell encapsulation with an emphasis on the fluid dynamics of droplet breakup and internal mixing as they directly influence microcapsule’s size and structure. We will then discuss two on-chip manipulation strategies: sorting and extraction from oil into aqueous phase, which can be integrated into droplet-based microfluidics and significantly improve the qualities of cell-laden hydrogel microcapsules. Finally, we will review various applications of hydrogel microencapsulation for 3D in vitro culture on cell growth and proliferation, stem cell differentiation, tissue development, and co-culture of different types of cells.

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Section: On-chip Manipulations For Cell-laden Hydrogel Microcapsulesmentioning

confidence: 99%

Generation and manipulation of hydrogel microcapsules by droplet-based microfluidics for mammalian cell culture

et al. 2017

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“…Despite the great advances in droplet‐based single‐cell isolation, it still remains challenging to ensure no more than one cell encapsulation. The traditional strategy is to use limited dilution based on Poisson distribution, which results in low occupancy of single cell with quantities of empty droplets . To maximize the percentage of single‐cell‐containing droplets, one alternation is to remove empty droplets with self‐sorting system .…”

Section: Single‐cell Isolationmentioning

confidence: 99%

Microfluidic Single‐Cell Omics Analysis

Wang

et al. 2019

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The commonly existing cellular heterogeneity plays a critical role in biological processes such as embryonic development, cell differentiation, and disease progress. Single‐cell omics‐based heterogeneous studies have great significance for identifying different cell populations, discovering new cell types, revealing informative cell features, and uncovering significant interrelationships between cells. Recently, microfluidics has evolved to be a powerful technology for single‐cell omics analysis due to its merits of throughput, sensitivity, and accuracy. Herein, the recent advances of microfluidic single‐cell omics analysis, including different microfluidic platform designs, lysis strategies, and omics analysis techniques, are reviewed. Representative applications of microfluidic single‐cell omics analysis in complex biological studies are then summarized. Finally, a few perspectives on the future challenges and development trends of microfluidic‐assisted single‐cell omics analysis are discussed.

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“…The last decade has yielded an exponential rise in new methods to analyze single cells 1,2 , revealing critical insights into cellular diversity [3][4][5][6][7] , tissue organization [8][9][10] , and organism development [11][12][13][14] . In particular, droplet microfluidics has emerged as a powerful class of single-cell isolation techniques due to its unprecedented scale (0.1 -10M droplets per run), throughput (0.1 -30 kHz generation rate), and efficiency ($0.10 -$0.50 per cell) [15][16][17] . Novel droplet assays have enabled thousands of single cells to be profiled by genome 18,19 , epigenome 20,21 , transcriptome 3,4,22 , or proteome [23][24][25] , leading to the generation of the first whole-organism cell atlases [26][27][28][29] .…”

Section: Introductionmentioning

confidence: 99%

Double Emulsion Picoreactors for High-Throughput Single-Cell Encapsulation and Phenotyping via FACS

Brower

Khariton

Suzuki

et al. 2020

Preprint

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In the past five years, droplet microfluidic techniques have unlocked new opportunities for the high-throughput genome-wide analysis of single cells, transforming our understanding of cellular diversity and function. However, the field lacks an accessible method to screen and sort droplets based on cellular phenotype upstream of genetic analysis, particularly for large and complex cells. To meet this need, we developed Dropception, a robust, easy-to-use workflow for precise single-cell encapsulation into picoliter-scale double emulsion droplets compatible with high-throughput phenotyping via fluorescence-activated cell sorting (FACS). We demonstrate the capabilities of this method by encapsulating five standardized mammalian cell lines of varying size and morphology as well as a heterogeneous cell mixture of a whole dissociated flatworm (5 -25 µm in diameter) within highly monodisperse double emulsions (35 µm in diameter). We optimize for preferential encapsulation of single cells with extremely low multiplecell loading events (<2% of cell-containing droplets), thereby allowing direct linkage of cellular phenotype to genotype. Across all cell lines, cell loading efficiency approaches the theoretical limit with no observable bias by cell size. FACS measurements reveal the ability to discriminate empty droplets from those containing cells with good agreement to single-cell occupancies quantified via microscopy, establishing robust droplet screening at single-cell resolution. High-throughput FACS phenotyping of cellular picoreactors has the potential to shift the landscape of single-cell droplet microfluidics by expanding the repertoire of current nucleic acid droplet assays to include functional screening. Supporting InformationSupplemental Information including extended methods, a full workflow protocol, and referenced supplemental figures (SI, PDF).

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Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis

Cited by 90 publications

References 93 publications

Generation and manipulation of hydrogel microcapsules by droplet-based microfluidics for mammalian cell culture

Generation and manipulation of hydrogel microcapsules by droplet-based microfluidics for mammalian cell culture

Microfluidic Single‐Cell Omics Analysis

Double Emulsion Picoreactors for High-Throughput Single-Cell Encapsulation and Phenotyping via FACS

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