Autonomous screening of C. elegans identifies genes implicated in synaptogenesis

Crane, Matthew M.; Stirman, Jeffrey N.; Ou, Chan-Yen; Kurshan, Peri T.; Rehg, James M.; Shen, Kang; Lu, Hang

doi:10.1038/nmeth.2141

Cited by 102 publications

(97 citation statements)

References 30 publications

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“…Microfluidics has been used for multiple applications in worm biology in recent years 10,11 . These applications include preparing and immobilizing worms for imaging and microsurgery 12 ; rapid control of changes in chemical environment for studies of the chemosensory system 13 ; trapping of worms for quantification of undulatory dynamics 14 ; as well as animal sorting 15 and screening 16 . In particular, several approaches to long-term imaging have been proposed [17][18][19] .…”

Section: Introductionmentioning

confidence: 99%

Durable spatiotemporal surveillance of Caenorhabditis elegans response to environmental cues

Kopito

Levine

2014

Lab Chip

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Animal response to changes in environmental cues is a complex dynamical process that occurs at diverse molecular and cellular levels. To gain a quantitative understanding of such processes, it is desirable to observe many individuals, subjected to repeatable and well defined environmental cues over long time periods. Here we present WormSpa, a microfluidic system where worms are individually confined in optimized chambers. We show that worms in WormSpa are neither stressed nor starved, and in particular exhibit pumping and egg-laying behaviors equivalent to those of freely behaving worms. We demonstrate the applicability of WormSpa for studying stress response and physiological processes. WormSpa is simple to make and easy to operate, and its design is modular, making it straightforward to incorporate available microfluidic technologies. We expect that WormSpa would open novel avenues of research, hitherto impossible or impractical.

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Section: Introductionmentioning

confidence: 99%

Durable spatiotemporal surveillance of Caenorhabditis elegans response to environmental cues

Kopito

Levine

2014

Lab Chip

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“…For instance, using microfluidics to streamline imaging and perform genetic screens has allowed reducing screening time by several orders of magnitude compared to traditional assays where worms are individually and manually picked. 8,[48][49][50] In a chip, thousands of worms can be successively loaded, imaged, phenotyped, and sorted. Generally, fluorescent reporters are used to discriminate worms of different phenotypes ( Fig.…”

Section: Genetic Screeningmentioning

confidence: 99%

“…To address this, technical solutions using suction, 51,52 mechanical restriction by a valve membrane, 53 channel constriction, 54 and CO 2 anesthesia 53 have been demonstrated; a common and reliable microfluidic method is to use valves to trap the worm in a region and use in situ temperature control to cool (to $4 C) and immobilize the worm. 8,[48][49][50] Cooling is fast, reversible, and effective. Compared to traditional immobilization methods that rely primarily on the use of anesthetic drugs, cooling-based immobilization is non-invasive, allows for recovering the worms after imaging and limits adverse developmental effects.…”

Section: Genetic Screeningmentioning

confidence: 99%

“…12 More recently in the automated process, novel mutants were identified based on predefined criteria; using machine-learning techniques, the power of using software algorithms can be pushed one step further where the sorting criteria are determined by the software without human intervention. 48,50 Overall, genetic screening in microfluidics has become a mature technology; further development is now focused on enhancing the ability to detect more subtle phenotypes 48 and discovering new biological mechanisms. …”

Section: Genetic Screeningmentioning

confidence: 99%

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A perspective on optical developments in microfluidic platforms for Caenorhabditis elegans research

Aubry

2014

Biomicrofluidics

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Microfluidics offers unique ways of handling and manipulating microorganisms, which has particularly benefited Caenorhabditis elegans research. Optics plays a major role in these microfluidic platforms, not only as a read-out for the biological systems of interest but also as a vehicle for applying perturbations to biological systems. Here, we describe different areas of research in C. elegans developmental biology and behavior neuroscience enabled by microfluidics combined with the optical components. In particular, we highlight the diversity of optical tools and methods in use and the strategies implemented in microfluidics to make the devices compatible with optical techniques. We also offer some thoughts on future challenges in adapting advancements in optics to microfluidic platforms.

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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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Autonomous screening of C. elegans identifies genes implicated in synaptogenesis

Cited by 102 publications

References 30 publications

Durable spatiotemporal surveillance of Caenorhabditis elegans response to environmental cues

Durable spatiotemporal surveillance of Caenorhabditis elegans response to environmental cues

A perspective on optical developments in microfluidic platforms for Caenorhabditis elegans research

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

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