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In JoVE (1)
- Microfluidic-based Electrotaxis for On-demand Quantitative Analysis of Caenorhabditis elegans' Locomotion
Other Publications (3)
Articles by Pouya Rezai in JoVE
Microfluidic-based Electrotaxis for On-demand Quantitative Analysis of Caenorhabditis elegans' Locomotion
Justin Tong1, Pouya Rezai2, Sangeena Salam1, P. Ravi Selvaganapathy2, Bhagwati P. Gupta1
1Department of Biology, McMaster University, 2Department of Mechanical Engineering, McMaster University
Other articles by Pouya Rezai on PubMed
Lab on a Chip. Jan, 2010 | Pubmed ID: 20066250
The nematode (worm) Caenorhabditis elegans is one of the most widely studied organisms for biomedical research. Currently, C. elegans assays are performed either on petri dishes, 96-well plates or using pneumatically controlled microfluidic devices. In this work, we demonstrate that the electric field can be used as a powerful stimulus to control movement of worms in a microfluidic environment. We found that this response (termed electrotaxis) is directional, fully penetrant and highly sensitive. The characterization of electrotaxis revealed that it is mediated by neuronal activity that varies with the age and size of animals. Although the speed of swimming is unaffected by changes in the electric field strength and direction, our results show that each developmental stage responds to a specific range of electric field with a specific speed. Finally, we provide evidence that the exposure to the electric field has no discernible effect on the ability of animals to survive and reproduce. Our method has potential in precisely controlling, directing, and transporting worms in an efficient and automated manner. This opens up significant possibilities for high-throughput screening of C. elegans for drug discovery and other applications.
Effect of Pulse Direct Current Signals on Electrotactic Movement of Nematodes Caenorhabditis Elegans and Caenorhabditis Briggsae
Biomicrofluidics. Dec, 2011 | Pubmed ID: 22232698
The nematodes (worms) Caenorhabditiselegans and Caenorhabditisbriggsae are well-known model organisms to study the basis of animal development and behaviour. Their sinusoidal pattern of movement is highly stereotypic and serves as a tool to monitor defects in neurons and muscles that control movement. Until recently, a simple yet robust method to initiate movement response on-demand did not exist. We have found that the electrical stimulation in a microfluidic channel, using constant DC electric field, induces movement (termed electrotaxis) that is instantaneous, precise, sensitive, and fully penetrant. We have further characterized this behaviour and, in this paper, demonstrate that electrotaxis can also be induced using a pulse DC electric signal. Worms responded to pulse DC signals with as low as 30% duty cycle by moving towards the negative electrode at the same speed as constant DC fields (average speed of C. elegansâ€‰=â€‰296â€‰Â±â€‰43â€‰Î¼m/s and C. briggsaeâ€‰=â€‰356â€‰Â±â€‰20â€‰Î¼m/s, for both constant and pulse DC electric fields with various frequencies). C. briggsae was found to be more sensitive to electric signals compared to C. elegans. We also investigated the turning response of worms to a change in the direction of constant and pulse DC signals. The response for constant DC signal was found to be instantaneous and similar for most worms. However, in the case of pulse DC signal, alterations in duty cycle affected the turning response time as well as the number of responding worms. Our findings show that pulse DC method allows quantitative measurement of response behaviour of worms and suggest that it could be used as a tool to study the neuronal basis of such a behaviour that is not observed under constant DC conditions.
Lab on a Chip. Apr, 2012 | Pubmed ID: 22460920
The nematode (worm) C. elegans is one of the widely studied animal model organisms in biology. It develops through 4 larval stages (L1-L4) in 2 to 3 days before becoming a young adult. Biological assays involving C. elegans frequently require a large number of animals that are appropriately staged and exhibit a similar behaviour. We have developed a new method to synchronize animals that relies on the electrotactic response (electric field-induced motion) of C. elegans to sort them in parallel based on their age, size and phenotype. By using local electric field traps in a microfluidic device, we can efficiently sort worms from a mixed culture in a semi-continuous flow manner (with a minimum throughput of 78 worms per minute per load-run) and obtain synchronized populations of animals. In addition to sorting larvae, our device can also distinguish between young and old adults efficiently. Unlike fluorescent based sorting systems that use active imaging based feedback, this method is passive and automatic and uses the innate behaviour of the worm. Considering that the entire procedure takes only a few minutes to run and is cost-effective, it promises to simplify and accelerate experiments requiring homogeneous cultures of worms as well as to facilitate isolation of mutants that have abnormal electrotaxis. More importantly, our method of isolating and separating worms using locomotion as a defining characteristic promises development of advanced microfluidics-based systems to study the neuronal basis of movement-related defects in worms and facilitate high-throughput chemical screening and drug discovery.