Droplet-based microfluidic system for individual Caenorhabditis elegans assay

Shi, Weiwei; Qin, Jianhua; Ye, Nannan; Lin, Baiquan

doi:10.1039/b808753a

Cited by 246 publications

(239 citation statements)

References 18 publications

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“…Actually, several existing droplet trapping and stabilization techniques could be applied to immobilize the droplets. 10,[26][27][28] But, the droplet stabilization was not the topic of the present work. Here we conducted the experiment on a vibration free table to maintain a static distribution during testing.…”

Section: Droplet Encapsulation Of Random Fast Moving Pathogenic mentioning

confidence: 97%

“…10,[26][27][28] In addition, some excellent droplet generation mechanisms can also be applied to simplify droplet formation process. 36 Lastly, by FIG.…”

Section: Discussionmentioning

confidence: 99%

“…5,6 Encapsulation of single biological cells into droplets has also been demonstrated successfully for parallel single cell analysis and drug screening. [7][8][9][10][11] It is noteworthy that these cells encapsulated using droplet approaches often have a low motility in media solutions. a) Author to whom correspondence should be addressed.…”

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confidence: 99%

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Microfluidic droplet encapsulation of highly motile single zoospores for phenotypic screening of an antioomycete chemical

et al. 2011

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Highly motile Phytophthora sojae (P. sojae) zoospores of an oomycete plant pathogen and antioomycete candidate chemicals were encapsulated into microdroplets. Random fast self-motion of P. sojae zoospores was overcome by choosing an appropriate flow rate for a zoospore suspension. To influence stochastic loading of zoospores into a microfluidic channel, a zoospore suspension was directly preloaded into a microtubing with a largely reduced inner diameter. A relatively high single zoospore encapsulation rate of 60.5% was achieved on a most trivial T-junction droplet generator platform, without involving any specially designed channel geometry. We speculated that spatial reduction in the diameter direction of microtubing added a degree of zoospore ordering in the longitudinal direction of microtubing and thus influenced positively to change the inherent limitation of stochastic encapsulation of zoospores. Comparative phenotypic study of a plant oomycete pathogen at a single zoospore level had not been achieved earlier.Phenotypic changes of zoospores responding to various chemical concentration conditions were measured in multiple droplets in parallel, providing a reliable data set and thus an improved statistic at a low chemical consumption. Since each droplet compartment contained a single zoospore, we were able to track the germinating history of individual zoospores without being interfered by other germinating zoospores, achieving a high spatial resolution. By adapting some existing droplet immobilization and concentration gradient generation techniques, the droplet approach could potentially lead to a medium-to-high throughput, reliable screening assay for chemicals against many other highly motile zoospores of pathogens.

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Section: Droplet Encapsulation Of Random Fast Moving Pathogenic mentioning

confidence: 97%

“…10,[26][27][28] In addition, some excellent droplet generation mechanisms can also be applied to simplify droplet formation process. 36 Lastly, by FIG.…”

Section: Discussionmentioning

confidence: 99%

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Microfluidic droplet encapsulation of highly motile single zoospores for phenotypic screening of an antioomycete chemical

et al. 2011

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“…[29][30][31][32][33][34][35][36][37][38][39][40][41] Several microfluidic devices have been reported for immobilizing worms in enclosed channels; 29,37,39,42 however, they do not allow a micropipette to access the worm body for injection. Two microfluidic devices have been developed to facilitate manual C. elegans microinjection.…”

Section: Introductionmentioning

confidence: 99%

A microfluidic device for automated, high-speed microinjection of Caenorhabditis elegans

2016

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The nematode worm Caenorhabditis elegans has been widely used as a model organism in biological studies because of its short and prolific life cycle, relatively simple body structure, significant genetic overlap with human, and facile/inexpensive cultivation. Microinjection, as an established and versatile tool for delivering liquid substances into cellular/organismal objects, plays an important role in C. elegans research. However, the conventional manual procedure of C. elegans microinjection is labor-intensive and time-consuming and thus hinders large-scale C. elegans studies involving microinjection of a large number of C. elegans on a daily basis. In this paper, we report a novel microfluidic device that enables, for the first time, fully automated, high-speed microinjection of C. elegans. The device is automatically regulated by on-chip pneumatic valves and allows rapid loading, immobilization, injection, and downstream sorting of single C. elegans. For demonstration, we performed microinjection experiments on 200 C. elegans worms and demonstrated an average injection speed of 6.6 worm/min (average worm handling time: 9.45 s/ worm) and a success rate of 77.5% (post-sorting success rate: 100%), both much higher than the performance of manual operation (speed: 1 worm/4 min and success rate: 30%). We conducted typical viability tests on the injected C. elegans and confirmed that the automated injection system does not impose significant adverse effect on the physiological condition of the injected C. elegans. We believe that the developed microfluidic device holds great potential to become a useful tool for facilitating high-throughput, large-scale worm biology research. V C 2016 AIP Publishing LLC.

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“…[9][10][11][12][13] Recently, it has also been investigated for the study of gene analysis 14,15 and DNA computation. 16 Here, we report a microfluidic DNA computing processor for gene expression analysis and corresponding gene drug synthesis, in which the DNA computing process performs Boolean logic operations.…”

Section: Introductionmentioning

confidence: 99%

A microfluidic DNA computing processor for gene expression analysis and gene drug synthesis

Qin

Lin

2009

Biomicrofluidics

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Boolean logic performs a logical operation on one or more logic input and produces a single logic output. Here, we describe a microfluidic DNA computing processor performing Boolean logic operations for gene expression analysis and gene drug synthesis. Multiple cancer-related genes were used as input molecules. Their expression levels were identified by interacting with the computing related DNA strands, which were designed according to the sequences of cancer-related genes and the suicide gene. When all the expressions of the cancer-related genes fit in with the diagnostic criteria, positive diagnosis would be confirmed and then a complete suicide gene ͑gene drug͒ could be synthesized as an output molecule. Microfluidic chip was employed as an effective platform to realize the computing process by integrating multistep biochemical reactions involving hybridization, displacement, denaturalization, and ligation. By combining the specific design of the computing related molecules and the integrated functions of the microfluidics, the microfluidic DNA computing processor is able to analyze the multiple gene expressions simultaneously and realize the corresponding gene drug synthesis with simplicity and fast speed, which demonstrates the potential of this platform for DNA computing in biomedical applications.

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Droplet-based microfluidic system for individual Caenorhabditis elegans assay

Cited by 246 publications

References 18 publications

Microfluidic droplet encapsulation of highly motile single zoospores for phenotypic screening of an antioomycete chemical

Microfluidic droplet encapsulation of highly motile single zoospores for phenotypic screening of an antioomycete chemical

A microfluidic device for automated, high-speed microinjection of Caenorhabditis elegans

A microfluidic DNA computing processor for gene expression analysis and gene drug synthesis

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