Lifespan-on-a-chip: microfluidic chambers for performing lifelong observation of C. elegans

Hulme, S. Elizabeth; Shevkoplyas, Sergey S.; McGuigan, Alison P.; Apfeld, Javier; Fontana, Walter; Whitesides, George M.

doi:10.1039/b919265d

Cited by 210 publications

(283 citation statements)

References 45 publications

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“…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] . Available devices however immobilize worms (either continuously or repeatedly) for imaging, thus perturbing the worms and evoking cellular and neuronal response.…”

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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“…Worm growth has also been studied using an array of chambers in which L4 worms were individually loaded to monitor development to adulthood [26], or using a device developed to observe collections of 30-40 adults worms [27]. A device with multiple observation chambers for adult worms was exploited to study chemical effects on worm behaviour [28] and oil-inwater emulsions were used to encapsulate individual worm embryos [29,30] or L1s [31] which could develop to adulthood.…”

Section: Introductionmentioning

confidence: 99%

A size threshold governsCaenorhabditis elegansdevelopmental progression

Uppaluri

Brangwynne

2015

Proc. R. Soc. B.

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The growth of organisms from humans to bacteria is affected by environmental conditions. However, mechanisms governing growth and size control are not well understood, particularly in the context of changes in food availability in developing multicellular organisms. Here, we use a novel microfluidic platform to study the impact of diet on the growth and development of the nematode Caenorhabditis elegans . This device allows us to observe individual worms throughout larval development, quantify their growth as well as pinpoint the moulting transitions marking successive developmental stages. Under conditions of low food availability, worms grow very slowly, but do not moult until they have achieved a threshold size. The time spent in larval stages can be extended by over an order of magnitude, in agreement with a simple threshold size model. Thus, a critical worm size appears to trigger developmental progression, and may contribute to prolonged lifespan under dietary restriction.

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“…In both groups, all the worms survived at least 8 days, which is consistent with the previously reported lifespan of wide-type N2 worms. 53 Therefore, we conclude that there is no evidence showing that the worm development was adversely affected by the automated robotic injection.…”

Section: Post-injection Viability Testmentioning

confidence: 92%

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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Lifespan-on-a-chip: microfluidic chambers for performing lifelong observation of C. elegans

Cited by 210 publications

References 45 publications

Durable spatiotemporal surveillance of Caenorhabditis elegans response to environmental cues

Durable spatiotemporal surveillance of Caenorhabditis elegans response to environmental cues

A size threshold governsCaenorhabditis elegansdevelopmental progression

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

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