Microfluidic Approaches for Manipulating, Imaging, and Screening C. elegans

Gupta, Bhagwati P.; Rezai, Pouya

doi:10.3390/mi7070123

Cited by 58 publications

(40 citation statements)

References 96 publications

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“…Worms that are sufficiently immobilized by anesthetics will maintain a straight body, and a cover slip can be used to gently slide over the worms until they lay on their lateral side. To optimize our methods further, microfluidic devices could be used to straighten the animals and ensure entire muscle fibres on one of the lateral sides can be visualized [80].…”

Section: Limitations Of the Current Studymentioning

confidence: 99%

Disruption of genes associated with Charcot-Marie-Tooth type 2 lead to common behavioural, cellular and molecular defects in Caenorhabditis elegans

et al. 2020

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Charcot-Marie-Tooth (CMT) disease is an inherited peripheral motor and sensory neuropathy. The disease is divided into demyelinating (CMT1) and axonal (CMT2) neuropathies, and although we have gained molecular information into the details of CMT1 pathology, much less is known about CMT2. Due to its clinical and genetic heterogeneity, coupled with a lack of animal models, common underlying mechanisms remain elusive. In order to gain an understanding of the normal function of genes associated with CMT2, and to draw direct comparisons between them, we have studied the behavioural, cellular and molecular consequences of mutating nine different genes in the nematode Caenorhabditis elegans (lin-41/ TRIM2, dyn-1/DNM2, unc-116/KIF5A, fzo-1/MFN2, osm-9/TRPV4, cua-1/ATP7A, hsp-25/ HSPB1, hint-1/HINT1, nep-2/MME). We show that C. elegans defective for these genes display debilitated movement in crawling and swimming assays. Severe morphological defects in cholinergic motors neurons are also evident in two of the mutants (dyn-1 and unc-116). Furthermore, we establish methods for quantifying muscle morphology and use these to demonstrate that loss of muscle structure occurs in the majority of mutants studied. Finally, using electrophysiological recordings of neuromuscular junction (NMJ) activity, we uncover reductions in spontaneous postsynaptic current frequency in lin-41, dyn-1, unc-116 and fzo-1 mutants. By comparing the consequences of mutating numerous CMT2-related genes, this study reveals common deficits in muscle structure and function, as well as NMJ signalling when these genes are disrupted.

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Section: Limitations Of the Current Studymentioning

confidence: 99%

Disruption of genes associated with Charcot-Marie-Tooth type 2 lead to common behavioural, cellular and molecular defects in Caenorhabditis elegans

et al. 2020

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“…1, 016102 (2017) multi-step fabrication and bonding techniques and may not be practical beyond two layers of membranes. 38,41,45 The cost of consumables in the lPADs is much lower compared to those in agarose plates or polymeric microfluidic devices. For example, using wax printing, it is possible to fabricate approximately 100 paper-based lPADs (4 cm 2 of device area) in an 8.5 in.…”

Section: -6mentioning

confidence: 99%

“…35,36 In the field of neurobiology, individual worms can be rapidly immobilized in microfluidic chambers where laser nanosurgery can be performed on their axons. 37 These and many more examples [38][39][40][41] show how microfluidics can be leveraged to physically or chemically manipulate worms' microcosm, along with large-scale genetic and behavioral screening to predict the role of underlying genes affecting an observed behavior.…”

Section: Introductionmentioning

confidence: 99%

“…Most of the worm chips described earlier necessitate the use of multi-layer device fabrication, sophisticated pumps and valve operations, and advanced programming skills to control and synchronize system operations. [37][38][39][40][41] The cost of chemicals and consumables in microfluidics can be expensive to sustain for repeated experiments when a regular inventory of devices is needed to test the biological hypotheses. Because of the above reasons, routine C. elegans experiments are often performed on Petri dishes and well plates that offer design simplicity, easy accessibility, experimental flexibility, standardized manufacturing, and disposability.…”

Section: Introductionmentioning

confidence: 99%

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Flexible and disposable paper- and plastic-based gel micropads for nematode handling, imaging, and chemical testing

et al. 2017

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Today, the area of point-of-care diagnostics is synonymous with paper microfluidics where cheap, disposable, and on-the-spot detection toolkits are being developed for a variety of chemical tests. In this work, we present a novel application of microfluidic paper-based analytical devices (μPADs) to study the behavior of a small model nematode, Caenorhabditis elegans. We describe schemes of μPAD fabrication on paper and plastic substrates where membranes are created in agarose and Pluronic gel. Methods are demonstrated for loading, visualizing, and transferring single and multiple nematodes. Using an anthelmintic drug, levamisole, we show that chemical testing on C. elegans is easily performed because of the open device structure. A custom program is written to automatically recognize individual worms on the μPADs and extract locomotion parameters in real-time. The combination of μPADs and the nematode tracking program provides a relatively low-cost, simple-to-fabricate imaging and screening assay (compared to standard agarose plates or polymeric microfluidic devices) for non-microfluidic, nematode laboratories.

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“…[18, 19] The study of nanomaterials in C. elegans and a diversity of organisms using a collaborative (multi-center) approach can facilitate a cost-effective yet robust evaluation of the risk posed by nanomaterials in the environment and humans. [20, 21]…”

Section: Introductionmentioning

confidence: 99%

Materials and toxicological approaches to study metal and metal-oxide nanoparticles in the model organism Caenorhabditis elegans

et al. 2017

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Understanding the in vivo fate and transport of nanoparticles (NPs) is challenging, but critical. We review recent studies of metal and metal oxide NPs using the model organism Caenorhabditis elegans, summarizing major findings to date. In a joint transdisciplinary effort, we highlight underutilized opportunities offered by powerful techniques lying at the intersection of mechanistic toxicology and materials science,. To this end, we firstly summarize the influence of exposure conditions (media, duration, C. elegans lifestage) and NP physicochemical properties (size, coating, composition) on the response of C. elegans to NP treatment. Next, we focus on the techniques employed to study NP entrance route, uptake, biodistribution and fate, emphasizing the potential of extending the toolkit available with novel and powerful techniques. Next, we review findings on several NP-induced biological responses, namely transport routes and altered molecular pathways, and illustrate the molecular biology and genetic strategies applied, critically reviewing their strengths and weaknesses. Finally, we advocate the incorporation of a set of minimal materials and toxicological science experiments that will permit meta-analysis and synthesis of multiple studies in the future. We believe this review will facilitate coordinated integration of both well-established and underutilized approaches in mechanistic toxicology and materials science by the nanomaterials research community.

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Microfluidic Approaches for Manipulating, Imaging, and Screening C. elegans

Cited by 58 publications

References 96 publications

Disruption of genes associated with Charcot-Marie-Tooth type 2 lead to common behavioural, cellular and molecular defects in Caenorhabditis elegans

Disruption of genes associated with Charcot-Marie-Tooth type 2 lead to common behavioural, cellular and molecular defects in Caenorhabditis elegans

Flexible and disposable paper- and plastic-based gel micropads for nematode handling, imaging, and chemical testing

Materials and toxicological approaches to study metal and metal-oxide nanoparticles in the model organism Caenorhabditis elegans

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