Detection and collection system of target single cell based on respiration activity

摘要

This paper describes the systematic technique for detection and collection of single cell based on respiration activity. The micro-arrayed optical oxygen sensor was used for parallel monitoring of single cell respiration activity. This sensor consists of Nafion film containing ruthenium complex for oxygen sensing, and micro well array prepared with carbon black doped polydimethyl-siloxane (PDMS). Physical dimensions of the micro well were 10 μm in diameter and 10–15 μm in depth. Fluorescence intensity of each well image was increased by the decrease of oxygen concentration in micro well. Furthermore, difference of cell activity (ratio of viable and dead cells) could be detected by the difference of fluorescence intensity. We have succeeded in a parallel monitoring for single cell respiration activity by using a microarrayed oxygen sensor. Target single cell collection based on oxygen sensing results was realized by using fluorescent inverted microscope equipped with an automatic single cell collection unit. The single cell in the target micro well was sucked with medium by precisely controlling nozzle position and the pressure inside nozzle. The mouse lymphocyte in the target well alone was successfully collected by combining these two technologies.