High-Speed Manipulation of Microobjects Using an Automated Two-Fingered Microhand for 3D Microassembly.

To assemble microobjects including biological cells quickly and precisely, a fully automated pick-and-place operation is applied. In micromanipulation in liquid, the challenges include strong adhesion forces and high dynamic viscosity. To solve these problems, a reliable manipulation system and special releasing techniques are indispensable. A microhand having dexterous motion is utilized to grasp an object stably, and an automated stage transports the object quickly. To detach the object adhered to one of the end effectors, two releasing methods-local stream and a dynamic releasing-are utilized. A system using vision-based techniques for the recognition of two fingertips and an object, as well automated releasing methods, can increase the manipulation speed to faster than 800 ms/sphere with a 100% success rate (N = 100). To extend this manipulation technique, 2D and 3D assembly that manipulates several objects is attained by compensating the positional error. Finally, we succeed in assembling 80-120 µm of microbeads and spheroids integrated by NIH3T3 cells.

Multifunctional Noncontact Micromanipulation Using Whirling Flow Generated by Vibrating a Single Piezo Actuator.

A noncontact method that can achieve immobilization, transportation, and rotation in the microscale is desired in biological micromanipulation. A multifunctional noncontact micromanipulation method is proposed here based on a vibration-generated whirling flow. Resonance of a cantilever structure is utilized to extend the straight vibration of a single piezo actuator to the 2D circular vibration of a micropipette. The circular vibration in fluids can generate the whirling flow featured with low pressure in the core area and flow velocity gradient. The low pressure can immobilize the objects nearby and transport them together with the micropipette, and the flow velocity gradient is utilized to form a torque to rotate the immobilized object. Experiments of the microbeads are conducted to evaluate the claimed functions and quantify the key parameters that influence the rotation velocity. The cell spheroid is immobilized and rotated for 3D observation, and by assessing the viability of the cells containing in the spheroid, the proposed method is proved noninvasive to living cells. Finally, another important application in operations of mouse egg cells is shown, which indicates that the proposed method is a potential valuable tool in biological micromanipulation.

Hydrodynamic Tweezers: Trapping and Transportation in Microscale Using Vortex Induced by Oscillation of a Single Piezoelectric Actuator.

The demand for a harmless noncontact trapping and transportation method in manipulation and measurement of biological micro objects waits to be met. In this paper, a novel micromanipulation method named “Hydrodynamic Tweezers” using the vortex induced by oscillating a single piezoelectric actuator is introduced. The piezoelectric actuator is set between a micropipette and a copper beam. Oscillating the actuator at a certain frequency causes the resonance of the copper beam and extend 1D straight oscillation of the piezoelectric actuator to 2D circular oscillation of the micropipette, which induces a micro vortex after putting the micropipette into fluid. The induced vortex featuring low pressure in its core area can trap the object nearby. A robotic micromanipulator is utilized to transport the trapped objects together with the micropipette. Experiments of trapping and transportation microbeads are carried out to characterize the key parameters. The results show that the trapping force can be controlled by adjusting peak-peak voltage of the sinusoidal voltage input into the piezoelectric actuator.

Piezo-actuated parallel mechanism for biological cell release at high speed.

In this paper, a dynamic releasing approach is proposed for high-speed biological cell manipulation. A compact parallel mechanism for grasping and releasing microobjects is used to generate controllable vibration to overcome the strong adhesion forces between the end effector and the manipulated object. To reach the required acceleration of the end effector, which is necessary for the detachment of the target object by overcoming adhesion forces, vibration in the end effector is generated by applying sinusoidal voltage to the PZT actuator of the parallel mechanism. For the necessary acceleration, we focus on the possible range of the frequency of the PZT-actuator-induced vibration, while minimizing the amplitude of the vibration (14 nm) to achieve precise positioning. The effect of the air and liquid environments on the required vibration frequency for successful release is investigated. For the first time, release results of microbeads and biological cells are compared. Release of the biological cells with 100 % success rate suggests that the proposed active release method is an appropriate solution for adhered biological cells during the release task.

Vision-based automated single-cell loading and supply system.

Automated continuous individual cell transfer is a critical step in single-cell applications using microfluidic devices. Cells must be aspirated gently from a buffer before transferring to operation zone so as not to artificially perturb their biostructures. Vision-based manipulation is a key technique for allowing nondestructive cell transportation. In this paper, we presented a design for an automated single-cell loading and supply system that can be integrated with complex microfluidic applications for examining or processing one cell at a time such as the current nuclear transplantation method. The aim of the system is to automatically transfer mammalian donor ( approximately 15 microm) or oocyte ( approximately 100 microm) cells one by one from a container to a polydimethylsiloxane (PDMS) microchannel and then transport them to other modules. The system consists of two main parts: a single-cell suction module, and a PDMS-based microfluidic chip controlled by an external pump. The desired number of vacuumed cells can be directed into the microfluidic chip and stored in a docking area. From the batch, they can be moved to next module by activating pneumatic pressure valves located on two sides of the chip. The entire mechanism is combined with monitoring systems that perform detection/tracking and control.