Microsystems for C. elegans Mechanics and Locomotion Study (2013)
Type of ContentTheses / Dissertations
Thesis DisciplineMechanical Engineering
Degree NameDoctor of Philosophy
PublisherUniversity of Canterbury. Mechanical Engineering
AuthorsJohari, Shazlinashow all
Studying animal mechanics is crucial in order to understand how signals in the neuromuscular system contribute to an organism’s behaviour and how force-sensing organs and sensory neurons interact. In particular, the connection between the nerves and the muscles responsible for the force generation in the neuromuscular system needs to be established. Knowledge of the locomotion forces can be beneficial for the development of therapies for muscle disorders, neurodegenerative and human genetic diseases, such as muscular dystrophy. The simplicity of the nematode Caenorhabditis elegans’ (C. elegans) nervous system, which is limited to 302 neurons, has made it an excellent model organism for studying animal mechanics which include mechanosensation and locomotion at the neuronal level. The advent of miniaturized force sensing devices has led to the proposal of various approaches for measuring C. elegans locomotion forces. However, these existing devices are relatively complex, involving complicated microfabrication procedures and are incapable of measuring forces exerted by C. elegans in motion. This thesis addresses these shortcomings by introducing a force sensor capable of continuously measuring the forces generated by C. elegans in motion. The system consists of a micropillar-based device made of polydimethylsiloxane (PDMS) only and a vision-based algorithm for resolving the worm force from the deflection of the cantilever-like pillars. The measured force is horizontal and equivalent to a point force acting at half of the pillar height. The microdevice, sub-pixel resolution for visual tracking of the deflection, and experimental technique form an integrated system for measuring dynamic forces of moving C. elegans with force resolution of 3.13 uN for worm body width of 100 um. A simple device fabrication process based on soft-lithography and a basic experimental setup, which only requires a stereo microscope with off-the-shelf digital camera mean that this method is accessible to most biological science laboratories. The results demonstrate that the proposed device is capable of quantifying multipoint forces of moving C. elegans rather than single-point forces for a worm sample. This allows one to simultaneously collect force data from up to eight measurements points on different worm body parts. This is a significant step forward as it enables researchers to explicitly quantify the relative difference in forces exerted by the worm’s different body segments during the worms’ movements. The device’s capability to determine multipoint forces during nematode motion can also generate meaningful data to compare forces associated with different worm body muscles, gaining new understanding on how these muscles function. The forces measured during locomotion in the micropillars could also be used to differentiate mutant phenotypes. Apart from locomotion forces, the device is also capable of conducting concurrent measurement of other locomotion parameters such as speed, body amplitude and wavelength, as well as undulation frequency. This additional information can be useful to further quantify phenotypic behaviour of C. elegans and deepen the understanding of the theory behind worm locomotion forces. The relationship between C. elegans locomotion forces and their environment has also been analyzed by variation of the pillar arrangement and spacing. The results indicate that the microstructured environment significantly affects the worm’s contraction force, locomotion speed and the undulation frequency. In addition, an alternative measurement technique was provided to measure worm forces on other substrates, such that worm locomotion behaviour in varying environments can be investigated further. The combination of the conventional measurement technique with the findings of worm locomotion on a glass substrate reported show promise for biological measurements and other sensing application such as tactile force. Additional functions of on-chip worm selection, sorting, and imaging have also been integrated with the device, rendering its potential to accommodate for high-throughput application of C. elegans force measurement and locomotion studies in the future. The primary contributions of this thesis are centered around four topics: the development of the PDMS micropillar array and its application to study C. elegans locomotion forces, the analysis of C. elegans muscular forces and locomotion patterns in microstructured environments, the investigation of the worm locomotion forces using different substrates and finally the integration of the PDMS micropillar with PDMS microvalve for on-chip worm selection and imaging. Although the results presented in this thesis focus on wild type C. elegans, the method can be easily applied to its mutants and other organisms.