Abstract
A droplet-based microfluidic device for long-term culture and longitudinal observation of Caenorhabditis elegans was presented. The worms were encapsulated in W/O droplets and controllably introduced into separate chambers, where the medium/oil or medium/air interface can be manipulated by a gravity-actuated technique, for lifelong culture. This device also contained clamp structures for reversibly immobilization of each worm separately, and thus we were able to track more details about each worm. Several phenotypes of worms were monitored from their fourth larva stage to death. The relationship between worm body length, stroke frequency and lifespan with this device was then studied. This device had a potential in various longitudinal researches on C. elegans including aging, drug screening, toxicity evaluation and etc.
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Consortium (The C. elegans sequencing consortium). Genome sequence of the nematode C. elegans: A platform for investigating biology. Science 282, 2012–2018 (1998).
Hulme, S.E. & Whitesides, G.M. Chemistry and the worm: Caenorhabditis elegans as a platform for integrating chemical and biological research. Angew. Chem. Int. Edit. 50, 4774–4807 (2011).
Wolff, S. & Dillin, A. The trifecta of aging in Caenorhabditis elegans. Exp. Gerontol. 41, 894–903 (2006).
Drake, J., Link, C.D. & Butterfield, D.A. Oxidative stress precedes fibrillar deposition of Alzheimer’s disease amyloid beta-peptide (1–42) in a transgenic Caenorhabditis elegans model. Neurobiol. Aging 24, 415–420 (2003).
Pinkston, J.M., Garigan, D., Hansen, M. & Kenyon, C. Mutation that increase the life span of C-elegans inhibit tumor growth. Science 313, 971–975 (2006).
Kaletta, T. & Hengartner, M.O. Finding function in novel targets: C-elegans as a model organism. Nat. Rev. Drug Discov. 5, 387–398 (2006).
Shi, W.W., Qin, J.H., Ye, N.N. & Lin, B.C. Dropletbased microfluidic system for individual Caenorhabditis elegans assay. Lab. Chip 8, 1432–1435 (2008).
Chronis, N., Zimmer, M. & Bargmann, C.I. Microfluidics for in vivo imaging of neuronal and behavioral activity in Caenorhabditis elegans. Nat. Methods 4, 727–731 (2007).
Arous, J.B., Tanizawa, Y., Rabinowitch, I., Chatenay, D. & Schafer, W.R. Automated imaging of neuronal activity in freely behaving Caenorhabditis elegans. J. Neurosci. Methods 187, 229–234 (2010).
Shi, W.W. et al. Droplet microfluidics for characterizing the neurotoxin-induced responses in individual Caenorhabditis elegans. Lab. Chip 10, 2855–2863 (2010).
Chung, K., Crane, M.M. & Lu, H. Automated on-chip rapid microscopy, phenotyping and sorting of C. elegans. Nat. Method 5, 637–643 (2008).
Lockery, S.R. et al. Artificial dirt: microfluidic substrates for nematode neurobiology and behavior. J. Neurophysiol. 99, 3136–3143 (2008).
Herndon, L.A. et al. Stochastic and genetic factors influence tissue-specific decline in ageing C-elegans. Nature 419, 808–814 (2002).
Johnston, J., Iser, W.B., Chow, D.K., Goldberg, I.G. & Wolkow, CA. Quantitative image analysis reveals distinct structural transitions during aging in Caenorhabditis elegans tissues. PLoS One 3, e2821 (2008).
Allen, P.B. et al. Single-synapse ablation and longterm imaging in live C. elegans. J. Neurosci. Methods 173, 20–26 (2008).
Jenifer, C.T. et al. Droplet-based microfluidic platforms for the encapsulation and screening of mammalian cells and multicellular organisms. Chem. Biol. 15, 427–437 (2008).
Hulme, S.E. et al. Lifespan-on-a-chip: microfluidic chambers for performing lifelong observation of C. elegans. Lab. Chip 10, 589–597 (2010).
Krajniak, J. & Lu, H. Long-term high-resolution imaging and culture of C. elegans in chip-gel hybrid microfluidic device for developmental studies. Lab. Chip 10, 1862–1868 (2010).
Chung, K. et al. Microfluidic chamber arrays for wholeorganism behavior-based chemical screening. Lab. Chip 11, 3689–3697 (2011).
Zhang, K. et al. Comprehensive two-dimensional manipulations of picoliter microfluidic droplets sampled from nanoliter samples. Anal. Chem. 83, 8029–8034 (2011).
Zhang, K. et al. On-demand microfluidic droplet manipulation using hydrophobic ferrofluid as a continuous-phase. Lab. Chip 11, 1271–1275 (2011).
Yamada, H. et al. Fabrication of gravity-driven microfluidic device. Rev. Sci. Instrum. 79, 124301 (2008).
Huh, D. et al. Gravity-driven microfluidic particle sorting device with hydrodynamic separation amplification. Anal. Chem. 79, 1369–1376 (2007).
Liu, J.J. et al. Using a circular groove surrounded inlet to generate monodisperse droplets inside a microfluidic chip in a gravity-driven manner. J. Micromech. Microeng. 18, 095014 (2008).
Zhang, K. et al. A gravity-actuated technique for flexible and portable microfluidic droplet manipulation. Microfluid. Nanofluid. 9, 995–1001 (2010).
Zhang, Z. et al. Microchemostat-microbial continuous culture in a polymer-based instrumented microbioreactor. Lab. Chip 6, 906–913 (2006).
He, T.X. et al. A modified microfluidic chip for fabrication of paclitaxel-loaded poly(L-lactic acid) microspheres. Microfluid Nanofluid 10, 1289–1298 (2011).
Weibel, D.B., DiLuzio, W.R. & Whitesides, G.M. Microfabrication meets microbiology. Nat. Rev. Micro- biol. 5, 209–218 (2007).
Xia, Y.N. & Whitesides, G.M. Soft lithography. Angew. Chem. Int. Edit. 37, 550–575 (1998).
Shook, D.R. & Johnson, T.E. Quantitative trait loci affecting survival and fertility-related traits in Caenorhabditis elegans show genotype-environment interactions, pleiotropy and epistasis. Genetics 153, 1233–1243 (1999).
Wang, D.Y. & Xing, X.J. Assessment of locomotion behavioral defects induced by acute toxicity from heavy metal exposure in nematode Caenorhabditis elegans. J. Environ. Sci. 20, 1132–1137 (2008).
Huang, C., Xiong, C.J. & Kornfled, K. Measurements of age-related changes of physiological processes that predict lifespan of Caenorhabditis elegans. P. Natl. Acad. Sci. USA 101, 8084–8089 (2004).
Stiernagle, T. Maintenance of C. elegans. WormBook, ed, The C. elegans Research Community, Worm-Book (2006).
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Ying, D., Zhang, K., Li, N. et al. A droplet-based microfluidic device for long-term culture and longitudinal observation of Caenorhabditis elegans . BioChip J 6, 197–205 (2012). https://doi.org/10.1007/s13206-012-6301-6
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DOI: https://doi.org/10.1007/s13206-012-6301-6