Deterministic trapping, encapsulation and retrieval of single-cells

Sauzade, Martin; Brouzés, Eric

doi:10.1039/c7lc00283a

Cited by 33 publications

(29 citation statements)

References 54 publications

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“…This ensures that each trap always contains a single particle. This design principle has been used to trap cells 40–42 , beads 43 , particles 44, 45 , and embryos 46, 47 in microfluidic devices. In this work, we synthesized microparticles of 100 µm diameter via stop flow lithography (SFL) 33–37 , with 10 5 particles/hr throughput, to demonstrate parking and isolation in our platform.…”

Section: Resultsmentioning

confidence: 99%

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Microparticle parking and isolation for highly sensitive microRNA detection

2017

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Isolating small objects, such as particles, cells, and molecules, in individual aqueous droplets is useful for chemical and biological assays. We have developed a simple microfluidic platform to immobilize (park) microparticles at defined locations, and isolate particles in monodisperse droplets surrounded by immiscible oil. While conventional methods can only achieve stochastic encapsulation of objects within larger droplets, our in situ method ensures that a single particle is entrapped in a similar-sized droplet, with ~95% yield for parking and isolation. This enables time-lapse studies of reactions in confined volumes and can be used to perform enzymatic amplification of a desired signal to improve the sensitivity of diagnostic assays. To demonstrate the utility of our technique, we perform highly sensitive, multiplexed microRNA detection by isolating encoded, functional hydrogel microparticles in small aqueous droplets. Non-fouling hydrogel microparticles are attractive for microRNA detection due to favorable capture kinetics. By encapsulating these particles in droplets and employing a generalizable enzyme amplification scheme, we demonstrate an order of magnitude improvement in detection sensitivity compared to a non-amplified assay.

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Section: Resultsmentioning

confidence: 99%

“…In addition, devices are fabricated by simple molding methods with inexpensive materials (PDMS). Thus, we envision that this universal platform can be used with other soft, biological entities 42 and find utility in diverse biomedical applications.…”

Section: Resultsmentioning

confidence: 99%

Microparticle parking and isolation for highly sensitive microRNA detection

2017

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“…The traps are designed such that they could trap single cells with high efficiencies, yet also allow for a smooth phase change. We followed resistance based design guidelines as reported for microfluidic serpentine trap designs for droplets [52][53][54] and cells [55][56][57] . The design element concerns the location of the traps relative to the main channel such that both single cells trapping, and phase change can occur.…”

Section: 1mentioning

confidence: 99%

“…lower deformability). [55,58] Following trapping of the cells is the encapsulation of a single cell inside a droplet using a phase change procedure. Popular passive single-cell encapsulation is known to be a procedure that follows Poisson statistics, generating droplets with none, one or more cells.…”

Section: 1mentioning

confidence: 99%

One cell, one drop, one click: hybrid microfluidic mammalian single-cell isolation

Samlali

Ahmadi

Quach

et al. 2020

Preprint

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The process of generating a stable knockout cell line is a complex process that can take several months to complete. In this work, we introduce a microfluidic method that is capable of isolating single cells, selecting successful edited clones, and expansion of these isoclones. Using a hybrid microfluidics method, droplets in channels can be individually addressed using a co-planar electrode system. In our hybrid microfluidic device, we show that we can trap single cells and subsequently encapsulate them on demand into pL-sized droplets. Furthermore, individual cells inside the droplet can be released from the traps or merged with other droplets by simply applying an electric potential to the electrodes that is actuated through a user interface. We use this high precision control to sort and to recover single isoclones to establish monoclonal cell lines, which is demonstrated with a heterozygous NCI-H1299 lung squamous cell population resulting from loss-of-function eGFP and RAF1 gene knock-out transfections.

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“…It is of great value in the use of single cell for three major reasons: (1) In aspect of the length scale, the diameter of microdroplet is controllable and can easily match the size of single cells (10–1000 µm). (2) Single‐cell encapsulation is easy to accomplish by a random encapsulation method or at very high efficiency by ingenious design of microchannel with a little time and materials consumption. (3) Acting as independent reactors, microdroplets are valuable in the analysis of extracellular products at single cell level because the extracellular products like proteins or metabolites are restricted in the droplet.…”

Section: Advanced Microfluidic Techniquesmentioning

confidence: 99%

Advances in Microfluidics Applied to Single Cell Operation

Zhu

Chu

Wang

2018

Biotechnology Journal

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The field of microbiology have traditionally been concerned with and focused on studies at the population level. Microfluidic platforms have emerged as important tools for biology research at a small scale, even down to a single cell level. The spatial and temporal control of cells and stimuli transported by microfluidic channels in well-designed microsystems realized the studies of specific cells in a controlled microenvironment. The true cellular physiology responses, which are obtained mostly by inference from population-level data, could be revealed in this way. Nowadays, significant applications like cell culture, analysis, sorting, genomics, and proteomics at the single cell level have been achieved in microfluidic chips. Highly integrated microfluidic systems with complete bio-analytic functions are also coming forth and of great promise for single cell related physiology, biomedical, and high throughput screening research. Herein, the leads of technologies applied to single cell operation are reviewed. Challenges and potentials of these works are also summarized, to highlight fields for further research.

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Deterministic trapping, encapsulation and retrieval of single-cells

Cited by 33 publications

References 54 publications

Microparticle parking and isolation for highly sensitive microRNA detection

Microparticle parking and isolation for highly sensitive microRNA detection

One cell, one drop, one click: hybrid microfluidic mammalian single-cell isolation

Advances in Microfluidics Applied to Single Cell Operation

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