Long survival observation of C. elegans inside Environmental SEM for nanomanipulations

This paper presents a novel technique for long survival observation of Caenorhabditis elegans (C. elegans) inside Environmental-SEM (E-SEM) for nanomanipulations. We have developed the nanorobotic manipulation system inside an Environmental-Scanning Electron Microscope (E-SEM) integrated optical microscope (OM) for nanobiomanipulation platform. Our platform provides a real-time high resolution image under water-contained condition in nano-meter high-magnification image. One of the difficulties on the E-SEM observation is to maintain the biological sample on survival condition in long time. In this paper, we propose a coating method on the C. elegans or sample substrate for maintain the survival condition to keep in wet-condition. The proposed technique is useful to operate the nanomanipulation on C. elegans body, for example nano-injection of functional micro-nano beads as demonstrated in this paper.