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.

We have developed a novel technique for injecting and cutting animal cells for an automatic cloning system. Animal cells that have been chemically softened are aspirated into a narrow teflon tube, which is then inserted into a microfluidic chip. The tube's tip is inserted into a microchannel and releases the aspirated cells in the microchannel. The microfluidic chip contains two orthogonal channels. One is a narrow microchannel that aspirates and fixes a cell; in the other channel, a fluid flows at high velocity for cutting the aspirated cell. The thickness of the aspiration channel can be adjusted to fix the aspirated cell. Half the volume of the cell is aspirated into the narrow microchannel and high-speed flow is then generated to cut the cell into two. In this report, the cutting method and the injection method are described. Then an imaging process is also investigated for an automatic cutting.

This paper presents image stabilization for microscopy using horizontal visual feedback control of the objective lens through a five-bar linkage and piezoelectric actuators, and its application to in vivo imaging. Even very small in vivo motion due to heartbeat and breathing makes microscopic observation difficult by blurring the microscope image or impossible by sending a region of interest out of view. In order to remove those unwanted effects of the motion, we have introduced motion-canceling robotic technologies into microscopy. Our image stabilization system through motion-canceling provides users with stabilized image sequences with respect to trembling of in vivo subjects. The developed image stabilization system, in term of robotics, corresponds to a visual feedback control system that consists of a robotic mechanism and a high-speed vision. A high-speed camera installed in the microscope detects the motion of the in vivo subject having topically applied fiducials. To virtually cancel this motion, we move the objective lens, synchronizing the motions of the subject and the lens to remove the relative motion between the two. As a result, we observe motion-free images to m. This technology is one of the very demanding technologies in biological research for in vivo observation with high resolution. In this paper, we verify the effectiveness of the developed system through in vivo experiments.