A microfluidic device for rapid screening of chemotaxis-defective Caenorhabditis elegans mutants

Yang, Jianping; Chen, Zuanguang; Yang, Fan; Wang, Shuping; Hou, Fenghua

doi:10.1007/s10544-012-9719-7

Cited by 18 publications

(17 citation statements)

References 49 publications

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“…In the context of neuroimaging, this generally involves the examination of structural, rather than functional, aspects of neuronal development. Recently developed techniques in high‐throughput and automated imaging have allowed the development of different types of studies, enabling both forward and reverse genetics, and potentially drug screens, on features and phenotypes difficult to detect previously . Here we review briefly these technological developments.…”

Section: Imaging Of Fluorescent Markersmentioning

confidence: 99%

“…Further, image‐processing algorithms can be used to reduce the variability introduced by human error and automatically examine the phenotype of interest and decide whether a worm is mutant of interest. Examples of success with the latter approach include the identification of synaptic mutants by variants of a single‐channel confinement device with cooling and the identification of chemotaxis mutants with a device capable of generating controlled gradients . In parallel, reverse genetics typically uses known genetic perturbations, for example, RNAi knock‐downs.…”

Section: Genetic Screens With High‐throughput Imagingmentioning

confidence: 99%

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Trends in high‐throughput and functional neuroimaging in Caenorhabditis elegans

Cho

Zhao

2017

WIREs Mechanisms of Disease

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The nervous system of Caenorhabditis elegans is an important model system for understanding the development and function of larger, more complex nervous systems. It is prized for its ease of handling, rapid life cycle, and stereotyped, well-cataloged development, with the development of all 302 neurons mapped all the way from zygote to adult. The combination of easy genetic manipulation and optical transparency of the worm allows for the direct imaging of its interior with fluorescent microscopy, without physically compromising the normal physiology of the animal itself. By expressing fluorescent markers, biologists study many developmental and cell biology questions in vivo; by expressing genetically encoded fluorescent calcium indicators within neurons, it is also possible to monitor their dynamic activity, answering questions about the structure and function of neural microcircuitry in the worm. However, to successfully image the worm it is necessary to overcome a number of experimental challenges. It is necessary to hold worms within the field of view, collect images efficiently and rapidly, and robustly analyze the data obtained. In recent years, a trend has developed toward imaging a large number of worms or neurons simultaneously, directly exploiting the unique properties of C. elegans to acquire data on a scale, which is not possible in other organisms. Doing this has required the development of new experimental tools, techniques, and data analytic approaches, all of which come together to open new perspectives on the field of neurobiology in C. elegans, and neuroscience in general. WIREs Syst Biol Med 2017, 9:e1376. doi: 10.1002/wsbm.1376 For further resources related to this article, please visit the WIREs website.

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Section: Imaging Of Fluorescent Markersmentioning

confidence: 99%

Section: Genetic Screens With High‐throughput Imagingmentioning

confidence: 99%

Trends in high‐throughput and functional neuroimaging in Caenorhabditis elegans

Cho

Zhao

2017

WIREs Mechanisms of Disease

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“…When considering high-throughput manipulation, automatic classification of worms (e.g., wild-type from mutants) becomes of high relevance. Typically, sorting involves individual C. elegans loading and separation, for instance, according to genetic phenotype for downstream analysis [31,32,34,36,[56][57][58][59][60]. Together with real-time rapid image extraction and data processing, media flow in the microfluidic channel is driven by a syringe and is controlled by on-chip functional components, such as PDMS valves.…”

Section: Microfluidic Techniques For C Elegans Manipulationmentioning

confidence: 99%

“…One application where microfluidics and fluorescent-based imaging open up aspects that would remain hidden from traditional laboratory techniques is drug screening. C. elegans can be an effective test-bed for a wide range of water-soluble chemical compounds (e.g., glycerol [30,66,74,75], anticancer drugs [48], heavy metals [54], sodium chloride NaCl [58,65,71,83,111,112], copper(II) chloride CuCl 2 [66,74], levamisole [70], manganese [102], antibiotics [104], isoamyl alcohol [113], cyanide [114], etc.). Microfluidic network manipulation allows the automation in a high-throughput manner and under reproducible experimental conditions while analysis of the nematode's chemosensitivity provides a detailed model of how neurons function together to generate behavioral response.…”

Section: Neuronal Studiesmentioning

confidence: 99%

Advanced Microfluidic Assays for Caenorhabditis elegans

Bakhtina¹,

MacKinnon²,

Korvink³

2016

Advances in Microfluidics - New Applications in Biology, Energy, and Materials Sciences

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The in vivo analysis of a model organism, such as the nematode Caenorhabditis elegans, enables fundamental biomedical studies, including development, genetics, and neurobiology. In recent years, microfluidics technology has emerged as an attractive and enabling tool for the study of the multicellular organism. "dvances in the application of microfluidics to C. elegans assays facilitate the manipulation of nematodes in high-throughput format and allow for the precise spatial and temporal control of their environment. In this chapter, we aim to illustrate the current microfluidic approaches for the investigation of behavior and neurobiology in C. elegans and discuss the trends of future development.Keywords: C. elegans, chip-based, manipulation, microfluidics, model organism . IntroductionThe invertebrate Caenorhabditis elegans, Drosophila melanogaster, and the vertebrate zebrafish Danio rerio are the most widely studied multicellular organisms. The in vivo analysis of these model organisms allows the understanding of many complex physiological processes, addressing many of the questions relevant to human biology. The choice of model organism depends on the biological question under investigation. For example, C. elegans is simple enough to be experimentally tractable. It has a short life cycle days at °C and lifespan days at °C , passes through four larva L L stages and an adult stage [ ]. Its small size . mm long and µm wide , transparent body at all life stages, and preferred food source Escherichia coli simplify its maintenance on agar plates or liquid cultures allowing visualization of individual cells and organs in intact animals. C. elegans possesses one of the simplest central nervous systems the adult hermaphrodite has neurons . "ecause it is so well studied, rapid identification of signaling pathways, for instance, in studies of aging, has © 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. become possible. "bout % of human disease genes have an orthologue in the genome of C. elegans, including those genes associated with "lzheimer disease, Parkinson's disease, Huntington's disease HD , and many other neurodegenerative disorders [ ]. This astonishing degree of correspondence permits the modeling of human ailments in a simple invertebrate without involving actual human subjects and provides a meaningful insight into the pathogenesis of a complex disease phenotype.Traditionally, behavioral genetics is employed as a prime method for neurobiological studies in C. elegans. It is based on manual worm manipulation on a Petri dish or a multiwell plate, and monitoring the effects on various biological processes, such as growth and fertility, by visual inspection. Refreshment of old buffer solutions by a fresh solution is invasive and causes stress both to the lar...

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“…Unfortunately, the worm in this non-physiological semi-restrained environment is not preferred for chemotaxis behavior studies. Recently, static diffusion-based microfluidic methods were established for generating chemical gradients to investigate the chemotaxis of C. elegans and to screen chemotaxis-defective mutants [26,27]. However, these methods need to take a longer time to form a chemical gradient compared to fluid flow-based methods.…”

Section: Introductionmentioning

confidence: 99%

Quantitative analysis of Caenorhabditis elegans chemotaxis using a microfluidic device

Tan

et al. 2015

Analytica Chimica Acta

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A microfluidic device for rapid screening of chemotaxis-defective Caenorhabditis elegans mutants

Cited by 18 publications

References 49 publications

Trends in high‐throughput and functional neuroimaging in Caenorhabditis elegans

Trends in high‐throughput and functional neuroimaging in Caenorhabditis elegans

Advanced Microfluidic Assays for Caenorhabditis elegans

Quantitative analysis of Caenorhabditis elegans chemotaxis using a microfluidic device

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