A simple culture system for long-term imaging of individual C. elegans

Pittman, Will Emmett; Sinha, Drew B; Zhang, William B.; Kinser, Holly E; Pincus, Zachary

doi:10.1039/c7lc00916j

Cited by 26 publications

(19 citation statements)

References 56 publications

(75 reference statements)

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“…In worms, midline tracking can be done in real time for dozens of individuals 26 . Combining multiworm midline tracking with custom sample chambers 34,38 would enable cradle-to-grave tracking of an animal's complete movements with sub-second resolution, providing valuable information about behavioural ageing and healthspan that could be combined with genetic and drug perturbation and compared with other physiological changes. In mice, the highest resolution tracking is typically done in for limited periods in fairly featureless 'open fields' 39,40 , but longer timescales can be accessible in systems that track mice in their home cages [41][42][43][44] .…”

Section: Bridging Behavioural Timescalesmentioning

confidence: 99%

Ethology as a physical science

2018

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Behaviour is the ultimate output of an animal's nervous system and choosing the right action at the right time can be critical for survival. A quantitative understanding of behaviour would therefore advance research in neuroscience as well as ecology and evolution. However, animal posture typically has many degrees of freedom and behavioural dynamics vary on timescales ranging from milliseconds to years, presenting both technical and conceptual challenges. Here we review 1) advances in imaging and computer vision that are making it possible to capture increasingly complete records of animal motion and 2) new approaches to understanding the resulting behavioural data sets. With the right analytical approaches, these data are allowing researchers to revisit longstanding questions about the structure and organisation of animal behaviour and to put unifying principles on a quantitative footing. Contributions from both experimentalists and theorists are leading to the emergence of a physics of behaviour and the prospect of discovering laws and developing theories with broad applicability. We believe that there now exists an opportunity to develop theories of behaviour which can be tested using these data sets leading to a deeper understanding of how and why animals behave.

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Section: Bridging Behavioural Timescalesmentioning

confidence: 99%

Ethology as a physical science

2018

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“…For example, one method consists of agarose hydrogel compartments, which allows parallel monitoring at high optical resolution of up to 25 animals simultaneously 34 and throughout larval development 35 . A second method constrains animals on a polyethylene glycol hydrogel substrate to individual "corrals", and allows longitudinal imaging of several individuals from embryo to aged adult 36 . In addition, a microfluidics-based method entails growing the worms in liquid microdroplets, but in this device, longevity is severely compromised and behavior is different from that seen on solid media 37 .…”

Section: Comparison With Other Methodsmentioning

confidence: 99%

Quantitative imaging of sleep behavior in Caenorhabditis elegans and larval Drosophila melanogaster

et al. 2019

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Sleep is nearly universal among animals, yet remains poorly understood. Recent work has leveraged simple model organisms, such as Caenorhabditis elegans and Drosophila melanogaster larvae, to investigate the genetic and neural bases of sleep. However, manual methods of recording sleep behavior in these systems are labor intensive and low in throughput. To address these limitations, we developed methods for quantitative imaging of individual animals cultivated in

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“…To reduce the need for manual transfers, many labs utilize a strong progeny-blocking drug (2’-deoxy-5-fluorouridine, FUdR) to maintain an adult-only population 14–16 . An alternative to this approach is the use of sterile mutants 17–19 .…”

Section: Introductionmentioning

confidence: 99%

NemaLife: A structured microfluidic culture device optimized for aging studies in crawlingC. elegans

Rahman

Edwards

Birze

et al. 2019

Preprint

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13Electronic Supplementary Information (ESI) available: Movies demonstrating progeny washing. Worm 14 development and arena optimization information. Protocol for lifespan assay in a microfluidic device. Abstract 34Caenorhabditis elegans is a powerful animal model in aging research. Standard longevity assays 35 on agar plates involve the tedious task of picking and transferring animals to prevent younger 36 progeny from contaminating age-synchronized adult populations. Large-scale studies employ 37 progeny-blocking drugs or sterile mutants to avoid progeny contamination, but such 38 manipulations change adult physiology and alter the influence of reproduction on normal aging. 39Moreover, for some agar growth-based technology platforms, such as automated lifespan 40 machines, reagents such as food or drugs cannot be readily added/removed after initiation of 41 the study. Current microfluidic approaches are well-suited to address these limitations, but in 42 their liquid-based environments animals swim rather than crawl, introducing swim-induced 43 stress in the lifespan analysis. Here we report a simple microfluidic device that we call NemaLife 44 that features: 1) an optimized micropillar arena in which animals can crawl, 2) sieve channels 45 that separate progeny and prevent the loss of adults from the arena during culture 46 maintenance, and 3) ports which allow rapid accessibility to feed the adult-only population and 47 emulate the body gait, locomotion, and lifespan of animals reared on agar. We validated our 51 approach with longevity analyses of classical aging mutants (daf-2, age-1, eat-2, and daf-16) 52 and animals subjected to RNAi knockdown of age-related genes (age-1 and daf-16). We also 53 showed that healthspan measures such as pharyngeal pumping and tap-induced stimulated 54 reversals can be scored across the lifespan. Overall, the capacity to generate reliable lifespan 55 and physiological data from the NemaLife chip underscores the potential of this device to 56 accelerate healthspan and lifespan investigations in C. elegans. 57 58 59 60 61 62 63 64 65Aging is a significant risk factor for a broad range of diseases including neurodegenerative 66 disorders, diabetes and cancer 1-5 . With the growing aging population, the socioeconomic 67 burden attributed with age-associated diseases is staggering and development of therapies that 68 promote healthy aging is imperative. C. elegans is a powerful model organism for aging 69 investigations with a short lifespan (3-5 weeks), remarkable genetic similarity with humans (~ 70 38 % orthologs 6 ) and conserved signaling pathways 7 . Additionally, a fully mapped genome 8 and 71 incredible genetic plasticity 9,10 makes C. elegans an attractive tool for aging studies. Advances in 72 fluorescent microscopy 11 and genomic technology (RNAi, CRISPR) 12,13 have further expanded 73 the number of possible ways in which C. elegans can be used to study healthy aging. 74Lifespan analysis has become a classic method for evaluating the effects of a wide variety of 75 gene...

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A simple culture system for long-term imaging of individual C. elegans

Cited by 26 publications

References 56 publications

Ethology as a physical science

Ethology as a physical science

Quantitative imaging of sleep behavior in Caenorhabditis elegans and larval Drosophila melanogaster

NemaLife: A structured microfluidic culture device optimized for aging studies in crawlingC. elegans

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