JPH0675170A - Manipulator - Google Patents

Abstract

PURPOSE:To efficiently carry a cell without destroying and to capture it with non-contact. CONSTITUTION:This manipulator is equipped with at least a pair of diaphragms 22, 23 confronting with each other in a parallel state and arranged so as to be positioned in liquid 12 including the cell 1, ultrasonic oscillators 32, 33 which generate the ultrasonic oscillation of the diaphragms by connecting to the diaphragms 22, 23, respectively, and a driving circuit which applies a sine wave voltage provided with prescribed frequency and phase difference to each of the ultrasonic oscillators 32, 33. Ultrasonic waves radiated from the pair of diaphragms 22, 23 to the liquid 12 are controlled so that the wavelength of 1/4 of odd-number times or even-number times can coincide with the 1/2 of distance between the diaphragms 22, 23 and also, phase difference of 180 deg. or 0 deg. can be obtained, and the node or loop of the standing wave of the ultrasonic wave can be generated selectively at a midpoint between the diaphragms, and the cell can be captured or eliminated at a midpoint.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a manipulator for capturing an object floating in a liquid, and more particularly to a manipulator used for cell manipulation performed under an optical microscope.

[0002]

2. Description of the Related Art FIG. 4 shows a state in which cell manipulation for gene recombination is performed under an optical microscope. In the figure, 2 is a micromanipulator, and the suction means (not shown) exerts a negative pressure on the fluid inside the tube to fix the cells 1 in the suction state. Reference numeral 3 denotes a micropipette which is inserted into the cell 1 from a position facing the micromanipulator 2, and the content is injected into the cell 1 by a micro-discharging device (not shown) when the cell 1 is inserted. Therefore, in the actual operation of cells, it is necessary to position the micromanipulator 2 and the micropipette 3 precisely.

FIG. 5 shows a positioning device for performing such positioning on the micromanipulator 2. In that case,
Since the positioning device of the micropipette 3 has the same structure, this description is omitted. Micro manipulator 2
Is rigidly connected to a piston 5 that slides in the first hydraulic cylinder 4. The hydraulic oil 6 filling the inside of the first hydraulic cylinder 4 passes through the pipe 7 to the second hydraulic cylinder 8
It is connected to the. A piston 9 is slidably provided inside the second hydraulic cylinder 8. This piston 9 is connected to a rotatable lever 10, and this lever 1
When 0 is operated by hand, the piston 9 slides back and forth. The cross-sectional area of the piston 9 in the second hydraulic cylinder 8 is the first
Is smaller than the cross-sectional area of the piston 5 of the hydraulic cylinder 4, the large movement of the piston 9 is converted into a fine movement in the piston 5, and the fine positioning of the micromanipulator 2 connected to the piston 5 is performed. Is possible. In FIG. 5, 11 is a suction means for adsorbing and fixing the cells 1 to the micromanipulator 2.

[0004]

As described above, the positioning device for the micromanipulator and the micropillet is designed only for minute positioning, and has a small movable range and a slow moving speed. ing. However, in the actual operation of cells, as shown in FIG. 6, it is necessary to sequentially capture a plurality of cells 1 floating in the same liquid 12 or to move cells 1 apart from the work flow area to the work area. However, in such a case, it was extremely troublesome to move the micromanipulator to capture the target cells. Further, since the micromanipulator mechanically contacts with cells to aspirate, depending on the type of cells, while being moved in the liquid 12, the adsorbed portion of the cells may be damaged due to the resistance of the surrounding liquid 12, or the cells may be damaged. There was a problem that the entire shape was distorted.

The present invention has been made in consideration of the above circumstances, and provides a manipulator capable of capturing a target cell in a non-contact manner and not damaging the cell even when it travels a long distance. The purpose is to

[0006]

The manipulator of the present invention is used when the cells are moved under an optical microscope, and the micromanipulator shown in FIG. 5 is used together in the cell manipulation after the movement. . Therefore, the manipulator of the present invention, at least a pair of diaphragms arranged so as to be positioned in the liquid containing the observation object and facing each other in parallel with each other, and the diaphragms connected to the respective diaphragms An ultrasonic oscillator for ultrasonically vibrating and a drive circuit for applying a sinusoidal voltage having a predetermined frequency and a phase difference to each ultrasonic oscillator are provided, and the ultrasonic wave is radiated into the liquid from the pair of diaphragms. For ultrasonic waves, 1/4 of the wavelength is an odd multiple or even multiple of 1 /
It is controlled to have a phase difference of 180 ° or 0 °, which is equal to 2, and selectively causes a node or antinode of the standing wave of ultrasonic waves to be generated at the midpoint between the diaphragms, and the cell at this midpoint. It is characterized by capturing or eliminating.

[0007]

1 to 3 show an embodiment of the present invention.
In FIG. 1, a part of a rigid rectangular frame body 20 is extended, and the extended portion 20 a is an XY table 2.
The entire frame body 20 can be positioned in a two-dimensional plane by being connected to positioning means such as 1. 4 inside the frame 20
Vibration plates 22, 23, 24, 25 are provided on the side surfaces. The vibrating plates 22, 23, 24, 25 are attached to the frame body 20 via ultrasonic transducers 32, 33, 34, 35, respectively.
When a voltage is applied to each of the diaphragms 22, 23, 24, 2
The ultrasonic waves from the ultrasonic vibrations 32, 33, 34, and 35 are propagated in 5 to radiate the ultrasonic waves. The vibrating plates 22, 23, 24, 25 are immersed in the liquid 12 in which the cells 1 dropped on the sample stage (not shown) of the optical microscope are suspended. Here, the diaphragms 22 and 23 at the facing position form a pair, and similarly, the diaphragms 24 and 25 at the facing position form a pair. As a result, the diaphragms 22, 23 and 24, 25 that form a pair are They are arranged in the orthogonal direction.

A driving circuit is connected to the ultrasonic transducers 32, 33, 34 and 35 to which the respective vibrating plates 22, 23, 24 and 25 are attached, and the ultrasonic transducers 3 forming a pair are connected.
A sine wave voltage having a predetermined frequency and phase difference is applied to 2, 33 and 34, 35. FIG. 2 shows a state where the drive circuits 42 and 43 are connected to the ultrasonic transducers 32 and 33 of the one pair of diaphragms 22 and 23, and although not shown, the other pair of diaphragms 24 and 25 ultrasonic transducers 3
A drive circuit is similarly connected to 4, 35.

In FIG. 1, 2 is a micromanipulator and 3 is a micropipette, which has the same structure as in FIG. The manipulator of the present embodiment is used for moving the micromanipulator 2 and the micropipette 3 in order to bring a target cell close thereto.

Next, the operation of this embodiment will be described with reference to FIGS. The sinusoidal voltage shown in FIG. 2 is applied from the drive circuits 42 and 43 to the pair of ultrasonic transducers 32 and 33, respectively. The frequency of this sine wave voltage is controlled so that 1/2 of the wavelength of the ultrasonic wave radiated into the liquid due to the vibration matches the distance D between the pair of diaphragms 22 and 23. . Then, the phases of the sine wave voltages are set to 180 degrees relative to each other.
° It is controlled so as to shift, and as shown in the figure,
A standing wave of ultrasonic waves is generated in the liquid 12 between the pair of vibrating plates 22 and 23. At this time, a vibration node a appears at the midpoint of the pair of diaphragms 22 and 23. In such a vibration node, the sound pressure is minimized, so that solid matter such as cells 1 in the liquid can be captured in the vibration node a.

In this state, in this embodiment, since two pairs of diaphragms are arranged in the orthogonal direction, it is possible to form an intersection of vibration nodes at the intersection of the axes from each pair of diaphragms.
As a result, the target cell 1 can be reliably captured at this intersection. Then, in this state, the frame body 20 is moved by the positioning means such as the XY table 21, so that the cells can be transferred to the tip portion of the micromanipulator 2 without contacting them.

FIG. 3 shows a pair of vibrators 22 and 23,
It is equal to the distance D between the diaphragms 22 and 23, and the phase difference is 0 °.
Shows a state in which a sinusoidal voltage that has become is applied. By applying this voltage, a standing wave is generated in the liquid as shown in FIG. An antinode b of the vibration is located at the midpoint of the vibration plates 22 and 23. Since the sound pressure is maximum in the antinode b portion of this vibration, the cells 1 floating in the liquid are excluded from the midpoint portion. Generally, in cell manipulation, when the micropipette 3 is inserted into a cell, other cells and foreign substances come close to each other, which hinders cell manipulation, but a standing wave as shown in FIG. 3 is generated in the liquid. By doing so, these can be eliminated.

Therefore, in the present embodiment as described above, cells scattered in a wide range in the liquid can be efficiently transferred, and the cells are trapped without contact, so that the cells can be collected before the cell manipulation. No damage. In addition, when cells are manipulated, it is possible to prevent other cells and foreign substances from entering the work area and disturbing the work by generating antinodes of standing waves in the work area.

[0014]

INDUSTRIAL APPLICABILITY As described above, according to the present invention, cells can be efficiently transferred from a wide range in a liquid to a working area during cell manipulation, and the cells are trapped in a non-contact manner to damage the cells. Without this, cell manipulation can be performed extremely quickly and reliably.

[Brief description of drawings]

FIG. 1 is a plan view of an embodiment of the present invention.

FIG. 2 is a side view showing the operation of cell trapping.

FIG. 3 is a side view showing an operation of cell exclusion.

FIG. 4 is a cross-sectional view showing cell manipulation.

FIG. 5 is a sectional view showing a positioning mechanism for cell manipulation.

FIG. 6 is a cross-sectional view showing another example of cell manipulation.

[Explanation of symbols]

 1 Cell 12 Liquid 20 Frame 22 Vibration Plate 23 Vibration Plate 32 Ultrasonic Transducer 33 Ultrasonic Transducer

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Atsuhiro Tsuchiya 2-43-2 Hatagaya, Shibuya-ku, Tokyo Olympus Optical Co., Ltd. (72) Inventor Takuma Sato 2-43-2 Hatagaya, Shibuya-ku, Tokyo Olin Path Optical Industry Co., Ltd. (72) Inventor Tomoki Funakubo 2-43-2 Hatagaya, Shibuya-ku, Tokyo Olin Path Optical Industrial Co., Ltd. (72) Yugo Imai 2-43-2 Hatagaya, Shibuya-ku, Tokyo Olin Pass Optical Industry Co., Ltd. (72) Inventor Toshiharu Tsubata 2-43-2 Hatagaya, Shibuya-ku, Tokyo Olympus Optical Industry Co., Ltd.

Claims (3)

[Claims] 1. A manipulator for manipulating cells under an optical microscope, wherein at least a pair of vibrating plates are arranged so as to face each other in parallel with each other and are located in a liquid containing an observation target, and respective vibrations. An ultrasonic oscillator connected to the plate for ultrasonically vibrating the diaphragm, and a drive circuit for applying a sinusoidal voltage having a predetermined frequency and phase difference to each ultrasonic oscillator, and the pair of vibrations. The ultrasonic wave radiated from the plate into the liquid is 1/4 of its wavelength.
The odd-numbered or even-numbered number of the waves is equal to ½ of the distance between the diaphragms, and is controlled to have a phase difference of 180 ° or 0 ° so that the ultrasonic standing wave of the ultrasonic wave is generated at the midpoint between the diaphragms. A manipulator characterized by selectively producing a node or an abdomen and trapping or excluding cells at this midpoint. 2. A pair of diaphragms are arranged so that the directions of standing waves of ultrasonic waves emitted by the pair of diaphragms are different.
The manipulator according to claim 1, wherein the manipulators are arranged in pairs, and the nodes or antinodes of the standing waves from each pair of diaphragms are crossed with each other, and cells are captured or excluded at the crossing points. 3. The pair of diaphragms and the ultrasonic vibrator are attached to a rigid frame body, and the frame body is positionally movable. The manipulator described in Crab.