Microfluidic chip-based C. elegans microinjection system for investigating cell-cell communication in vivo

Abstract

The propagation of intercellular calcium wave (ICW) is essential for coordinating cellular activities in multicellular organisms. However, the limitations of existing analytical methods hamper the studies of this biological process in live animals. In this paper, we demonstrated for the first time a novel microfluidic system with an open chamber for on-chip microinjection of C. elegans and investigation of ICW propagations in vivo. Worms were long-term immobilized on the side wall of the open chamber by suction. Using an external micro-manipulator, localized chemical stimulation was delivered to single intestinal cells of the immobilized worms by microinjection. The calcium dynamics in the intestinal cells expressing Ca(2+) indicator YC2.12 was simultaneously monitored by fluorescence imaging. As a result, thapsigargin injection induced ICW was observed in the intestinal cells of C. elegans. Further analysis of the ICW propagation was realized in the presence of heparin (an inhibitor for IP3 receptor), which allowed us to investigate the mechanism underlying intercellular calcium signaling. We expect this novel microfluidic platform to be a useful tool for studying cell-cell communication in multicellular organisms in vivo.

Keywords: Calcium imaging; Intercellular calcium wave; Microfluidic chip; Microinjection.