Fluorescent monitoring using microfluidics chip and development of syringe pump for automation of enucleation to automate cloning

We have developed a novel technique for fluorescent monitoring of a bovine oocyte nucleus for an automatic cloning device. Animal cells that have been chemically softened by cytochalasin and stained with Hoechst dye are aspirated into a thin microchannel of a microfluidic chip and stretched thin, allowing the nucleus of the expanded oocyte to be monitored. Half the volume of the oocyte is aspirated into the thin microchannel and a high-velocity fluid flow is generated in the wide microchannel to bisect the oocyte. Then, the half-oocytes are monitored to determine which contains the nucleus. To control flow velocity with high accuracy and rapid response, we also developed a syringe pump that is small, has no backlash, and has highly-accurate volume control and a good response for automatic cutting. In this report, we describe the monitoring method and construction of the syringe pump.