New Modes of GPCR Signalling
New Modes of GPCR Signalling
New Modes of GPCR Signalling
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Microvavle Immobilization <strong>of</strong> Intact C. Elegans for in vivo Calcium<br />
Imaging <strong>of</strong> Neuronal Activities in Response to External Stimuli<br />
Jingjing Wang, Xiaojun Feng, Wei Du, Bi-Feng Liu*<br />
Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for<br />
Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory,<br />
Department <strong>of</strong> Systems Biology, College <strong>of</strong> Life Science and Technology, Huazhong<br />
University <strong>of</strong> Science and Technology, Wuhan 430074, People’s Republic <strong>of</strong> China.<br />
Aim To immobilize intact young adult C. elegans for imaging and explore neuronal<br />
activities in response to the changing environments. Methods We adopted a harmless<br />
and innocuous method to immobilize awake behaving C. elegans in a micr<strong>of</strong>luidic<br />
device for imaging in vivo neuronal responses to controllable drug stimuli. A<br />
comb-shaped micr<strong>of</strong>luidic valve was developed for the immobilization <strong>of</strong> living C.<br />
elegans with minimal influence on its activity for a long period <strong>of</strong> time. Results Our<br />
micr<strong>of</strong>luidic device achieved the complete immobilization effect <strong>of</strong> living C. elegans,<br />
similar to the glue-based method. In coupling to a single-drug delivery system, we<br />
investigated the neuronal response and adaptation <strong>of</strong> the amphid polymodal ASH<br />
sensory neurons to repeated rapid, sharp stimuli such as high concentrations <strong>of</strong> glycerol.<br />
With a multi-drug delivery system, we sequentially stimulated the same animal with<br />
three different chemical reagents, resulting in successful differentiation between<br />
mutants and wild type. Conclusion The PDMS comb-shaped microvalve demonstrates<br />
the complete immobilization <strong>of</strong> awake behaving C. elegans. Micr<strong>of</strong>luidic devices also<br />
provide micrometer-sized scales for rapid and sharp change in microenvironment to<br />
observe neuron responses to controllable drug stimuli. Thus, sensitive imaging <strong>of</strong><br />
neuronal activities can be conducted in vivo. We expect that micr<strong>of</strong>luidic chips can be<br />
used to study different aspects <strong>of</strong> nervous system in C. elegans.<br />
Key Words: C. elegans, micr<strong>of</strong>luidic, comb-shaped valve, neuronal activities<br />
*Corresponding author<br />
Email: bfliu@mail.hust.edu.cn Tel: 86-27-87792203
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