JP4062669B2 - Micro object suction holding device - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a minute object suction and holding apparatus used as an improved hydraulic micromanipulator or microinjector used in a laboratory or the like and used to place a minute object on a microscope preparation.
[0002]
[Prior art]
Conventionally, in order to observe a microscopic object having a diameter of about 100 to 200 microns and a weight of about 0.5 to 4 microgram in a laboratory, etc. Must be taken from a predetermined storage location and placed in a predetermined location where it can be observed with a microscope.
However, when this type of egg cell such as a human being or an animal is placed in a place where it can be observed with a microscope or the like, this type of egg cell is not only a minute object, but its surface is soft and has a slight external force. Since the surface is easily destroyed, when it is moved, the suction holding movement is carefully performed using a special instrument / device. Therefore, this kind of egg cell is usually moved by sucking and holding the egg cell using a jig called a hydraulic micromanipulator or microinjector and placing it in a required place.
[0003]
FIG. 4 is a diagram showing an outline of a microinjector used for sucking and holding such a small object conventionally used. In FIG. 4, 101 is a pipette tube for sucking and holding egg cells and the like. The pipette tube 101 is connected to the flexible tube 103 and the main body 104 in a state of being filled with a predetermined liquid (liquid paraffin). Consists of configuration. The pipette tube 101 and the metal tube 102 are connected to the microinjector main body 104. The microinjector main body 104 includes a dial injector 105. By rotating the dial 106 left and right, It is configured so that a small amount of the enclosed liquid (for example, liquid paraffin) can be sucked or discharged.
[0004]
Therefore, when trying to suck and hold an egg cell having a diameter of about 100 to 200 microns, for example, at the tip of the pipette tube 101, the tip opening of the pipette tube 101 is filled with liquid, and in that state, is in close contact with the egg cell. When 106 is rotated and a negative pressure is applied to the opening of the pipette tube 101, minute egg cells are sucked and held in the opening of the pipette tube by the negative pressure. And it moves to a microscope observation area | region etc. in the state hold | maintained by suction.
After moving to the microscopic observation region, the dial 106 is rotated in the opposite direction, and positive pressure is applied to the pipette tube tip opening, and the egg cell is pushed out from the pipette tube tip opening so as to separate the close contact. Then, the detached egg cell is placed in the microscope observation region to observe the egg cell.
[0005]
[Problems to be solved by the invention]
In this way, the egg cells that are minute and whose surface is easily destroyed are moved to the microscope observation region. The operation of the microinjector, particularly the operation of the dial 106, sucks and discharges a minute thing. However, even if the dial is rotated too much, the soft cell surface may be broken by negative pressure, and operation of the microinjector requires some skill. It was. And even a skilled person may cause the dial 106 to rotate too much at any time, causing the same egg cell surface destruction. In addition, since these devices generally use oil which is an internally sealed liquid, there is a drawback that not only the device but also its operation maintenance is complicated.
In addition, with conventional devices, the tube that fills the oil and transmits positive pressure and pulling pressure from the dial at hand to the glass tube is long, so if it touches the tube at any moment, the pulling pressure is released. There was a problem that the egg cells were separated.
SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide a minute object suction holding device that can reliably suck and hold egg cells and the like whose surface is easily broken without requiring skill and move them to a target location.
[0006]
In view of the above-described object, the minute object suction holding apparatus according to claim 1 of the present application has an opening for sucking a minute object having a tip of 100 to 200 microns, and the other end is formed in a T shape with a larger diameter than the tip opening. A heating part disposed in the vicinity of the branch sealing end , and an ultrasonic transmitter for measuring the hardness of the surface of the sucked egg cell and / or an internal stirring super A glass pipette tube in which a sound wave vibrator is disposed, and a power supply unit that heats the heating unit, by energizing and heating the heating unit for a predetermined time in advance, expand the internal air in the glass pipette tube, A positive pressure is generated at the tip, and in that state, an egg cell or a microcapsule of a human, animal, or the like is brought into close contact with the tip, and then the energization is cut to bring the glass pipette tube into a cooled state, and the internal air is discharged. Shrink The negative pressure is generated at the tip, characterized in that so as to suck and hold the egg cell or microcapsules.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
The present invention will be described based on a preferred embodiment.
[First Embodiment]
FIG. 1 is a schematic view showing an outline of a minute object suction / holding device according to a first embodiment of the present invention.
In the figure, reference numeral 1 denotes a micro object suction holding device main body, and reference numeral 2 denotes a suction glass pipette tube (glass tube) whose tip is opened narrowly, followed by a large-diameter portion having a diameter of 0.5 mm. Have Reference numeral 3 denotes a heating nichrome wire wound in a coil shape a predetermined number of times around the large-diameter portion. Further, 4 is a switch, 5 is a DC power supply, 6 is a servo controller, and 7 is a servo mechanism. The servo controller 6 and the servo mechanism 7 are devices for performing minute movement precisely, and the servo mechanism 7 is hindered by rotating the three-way rotation controller of the servo controller 6 of the operator. The glass pipette tube 2 can be precisely moved three-dimensionally from side to side, back and forth or up and down. Although the servo controller 6 and the servo mechanism 7 are not unique to the minute object suction / holding device, the minute object suction / holding device according to the present invention is indispensable because the minute movement needs to be precisely performed. .
[0008]
The minute object suction and holding device according to the present invention uses these servo controller 6 and servo mechanism 7, and further expands and contracts by heating the internal air of the glass pipette tube 2 that interferes therewith. By utilizing the expansion / contraction action of air, a fine object such as an egg cell is sucked and held at the tip of the glass pipette tube 2 or released.
[0009]
FIG. 2 shows an outline of the nichrome wire structure wound around the hollow glass pipette tube 2 and the large diameter portion of the root. FIG. 2 is a schematic view of the nichrome wire structure wound around the hollow glass pipette tube 2 and the large diameter portion of the root. As described above, reference numeral 2 in the drawing is a suction glass pipette tube (glass tube) having a tip opened at a diameter of 130 microns. In the present embodiment, the glass pipette tube 2 includes a Narishige company. A glass tube manufactured by G-1 is used, and following the thinnest portion, a continuous thick portion having a diameter of 0.5 mm is provided, and the length is 40 mm. Reference numeral 3 denotes a heating nichrome wire wound around the thick-diameter portion in a predetermined coil shape. The resistance of the nichrome wire has a resistance value of 25Ω as a whole. Reference numeral 8 denotes a variable resistor for current adjustment. In this embodiment, a variable resistor of 100Ω is used. Reference numeral 9 denotes a switch, and reference numeral 10 denotes a DC power source.
[0010]
The mechanism for sucking and holding egg cells and the like with the glass pipette tube 2 wound with the heating nichrome wire 3 according to the present embodiment and further releasing it is as follows. That is, with the electrical configuration based on the above-described combination, egg cells can be aspirated by applying DC 10 V from the DC power source 10 for about 1 second. The current flowing at this time was measured and found to be 0.2A. Therefore, the heat amount is Q ₁ = 0.24 × i × E × t = 0.24 × 0.2 × 10 × 1 = 0.48 cal. At that time, since air expansion of about 1 mm was observed in the tip thin tube portion, when calculating the expansion of air in the glass pipette tube 2 due to this heat supply, the tip diameter of the glass pipette tube 2 was 130 micrometers, The air expands about 1 mm by the heat supply.
[0011]
That is, ignoring the volume change of the thin tube portion, the total volume V of air in the glass pipette tube 2 is V = 0.5 × 0.5 × 3.14 × 40 = 31.4 mm ³ , The total volume is increased by the expansion amount ΔV = 0.13 × 0.13 × 3.14 × 1 = 0.053 mm ³ by the heat amount of 0.48 cal, and this ΔV pushes the capillary volume, The small egg cell is released to the tip of the thin tube by pressurizing the amount of extrusion or by reducing the power when the power is turned off and the large diameter portion of the glass pipette tube 2 is cooled and the air in this portion is reduced. Or, it will be sucked and held.
[0012]
Incidentally, the air in the glass pipette tube 2 was subjected to a temperature increase of 0.5 ° C. by applying the V0.2A for 1 second, so that Q ₂ = 0.00. The amount of heat of 24 × 1.205 × 10 ⁻⁶ × 31.4 × 0.5 = 0.45 × 10 ⁻⁵ cal (Q = specific heat × weight × temperature change) was generated. In this case, the heat transfer efficiency is Q ₂ / Q ₁ × 100 = 0.00095%. The calculation was performed assuming that the specific heat of air was 0.24 kcal / kg · ° C. and the density of air was 1.205 kg / m ³ (20 ° C. · atmospheric pressure).
[0013]
Thus, by applying a current of DC 10V × 0.2A for about 1 second to the nichrome wire (25Ω) arranged in the large-diameter portion of the glass pipette tube 2 composed of the 130-micron-diameter tip thin tube as described above, By inflating the internal air and then cutting off the application, the air in the portion is cooled, and thereby the minute amount of eggs such as humans and animals, microcapsules, etc. that are in close contact with the tip of the glass pipette tube 2 The object can be moved while sucking for a predetermined time without damaging the surface, and then the internal air is expanded again by turning on the power again, and the glass pipette tube is expanded by the expansion of the air. It was possible to generate a European pressure on the tip 2 and release the surface of the egg cell that was sucked and held, and place the egg cell there.
[0014]
[Second Embodiment]
The above uses a straight glass pipette tube 2 as the first embodiment, but this uses a T-shaped glass pipette tube as shown in FIG. In addition to the heated part, an ultrasonic transmitter is installed at the bottom of the straight line, and a tactile sensor that measures the hardness of the surface of eggs and the like sucked at the tip of the ultrasonic wave emitted from this ultrasonic transmitter is placed. It is something that can be done.
[0015]
FIG. 3 is a diagram showing an outline of a glass pipette tube and a wound nichrome wire portion of a second embodiment in which a T-shaped glass pipette tube is used and an ultrasonic transmitter for a tactile sensor is arranged on a straight bottom portion. The same parts as those shown in the first embodiment are indicated by the same reference numerals. In the figure, 2 and 3 are a glass pipette tube and a coiled nichrome wire, respectively, but in this embodiment, the glass pipette tube 2 itself is formed in a T-shape as shown in FIG. A nichrome wire is wound around the branched end, and this portion can be heated, and an ultrasonic transmitter 11 constituting a tactile sensor is provided at the straight end of the T-shaped glass pipette tube 2. Is placed and sealed. In addition, the code | symbol 12 is a sealing device which installs a transmitter in the figure.
[0016]
In the present embodiment, the glass pipette tube 2 to be used has three ends in a T shape, but the inside is connected in a hollow shape, and the T shape is the same as in the first embodiment. When the branch end of the glass pipette is heated, the internal air is expanded by the heating, and the internal air is pushed out from the tip of the glass pipette tube 2 by the expansion force. When the energization to the nichrome wire is terminated in this state, the heating is completed, the internal air is reduced by natural cooling, and a negative pressure is generated at the tip of the glass pipette tube. Due to the negative pressure, egg cells and the like are sucked and held at the tip. And in the state of sucking and holding the egg cells, etc., after moving to the required place, for example, the microscope observation region, by energizing the nichrome wire again, the part is heated, the internal air is expanded, Thereby, a positive pressure is generated at the tip of the glass pipette tube 2 to release egg cells and the like from the tip.
[0017]
In the minute object suction and holding device according to the second embodiment, the surface hardness of the egg cell or the like can be measured while being sucked and held, so that the surface can be measured without being arranged in a separate hardness measuring device or the like. Hardness can be measured, and as described above, there is no possibility of destroying the surface during movement. Furthermore, operability, maintainability, etc. are reduced from the supplementary device using conventional oil, and the operability and maintainability are improved at each stage, and the experimental work efficiency is improved.
In the second embodiment, the ultrasonic transmitter for a tactile sensor for measuring the surface hardness of an egg cell or the like is arranged on the straight bottom of the T-shaped glass pipette tube. Instead of the ultrasonic transmitter, or together with this ultrasonic sensor ultrasonic transmitter, an internal stirring ultrasonic transducer that disturbs the inside of the microcapsule at the tip of the glass pipette tube may be arranged. By arranging an internal stirring ultrasonic vibrator that disturbs the inside of the microcapsule, for example, it is possible to stir a very small amount of substance, such as an intracellular substance taken out of a single cell, This is particularly useful when it is necessary to induce a chemical reaction.
[0018]
【The invention's effect】
A minute object suction holding device according to the present invention is a glass pipette tube having an opening of about 130 microns at the tip, a large-diameter sealing part at the multi-end, and a heating part arranged in the vicinity of the sealing part. And a power supply unit that heats the heating unit, and energizes and heats the heating unit for a predetermined time in advance to expand the internal air in the glass pipette tube and generate a positive pressure at the tip. Then, after the egg cells of humans, animals, etc. are brought into close contact with the tip, the glass pipette tube is cooled by cutting off the energization, the internal air is reduced, and negative pressure is generated at the tip. Since the egg cells etc. are sucked and held, there is no need to adjust the hydraulic pressure, etc., compared to conventional devices that use hydraulic pressure, etc., so that the device can be configured easily, and its operation, maintenance, etc. are simple Can be done There is an effect that.
Furthermore, the use of this device solves the problem of contact with a long tube, releasing the attractive pressure to some beat and leaving the egg cell away, and it is desirable to improve the efficiency of the work.
[0019]
Furthermore, the positive pressure and negative pressure of the tip opening can be adjusted based only on the internal volume of the glass pipette tube, the tip opening, and the amount of heating heat. Can be easily adjusted, and it has an excellent effect that a highly expandable device can be obtained.
Further, by using a T-shaped glass pipette tube as shown in the second embodiment, the heating portion is not provided at the straight end of the pipette tube, and the tactile sensor is attached to the straight end. By providing an ultrasonic transmitter and other sensor means, it is very easy to measure the surface hardness, etc. while sucking and holding the tip, or to perform internal disturbances. It has the outstanding effect that it can be set as the micro object suction holding | maintenance apparatus which combines.
Furthermore, by irradiating the glass tube (not inside the glass tube) with ultrasonic waves from the internal disturbance ultrasonic vibrator while sucking and holding the egg cells, the tip egg cell or the microcapsule can be disturbed.
[Brief description of the drawings]
FIG. 1 is a schematic diagram showing an outline of a minute object suction holding device according to a first embodiment of the present invention;
FIG. 2 is a schematic view of a nichrome wire structure wound around a hollow glass pipette tube 2 and a thick part of the root diameter;
FIG. 3 is a schematic view of a glass pipette tube and a wound nichrome wire portion according to a second embodiment in which a T-shaped glass pipette tube is used and an ultrasonic transmitter for a tactile sensor is arranged on a straight bottom. Figure showing the
FIG. 4 is a diagram showing an outline of a micro-injector used for sucking and holding such a small object conventionally used.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Micro object suction holding device main body 2 ... Glass pipette pipe | tube 3 for suction ... Nichrome wire 4 for heating ... Switch 5 ... DC power supply 6 ... Servo controller 7 ... Servo mechanism 8 ... Variable resistor for current adjustment 9 ... Switch 10 ... DC power supply 11 ... Ultrasonic transmitter 12 ... Sealing device 101 ... Pipette tube 102 ... Metal tube 103 ...・ Flexible tube 104 ... Micro injector main body 105 ... Dial type injector 106 ... Dial

Claims (1)

The tip has an opening for sucking a small object of 100 to 200 microns, and the other end has a large-diameter sealing portion formed in a T shape with a larger diameter than the tip opening , and is arranged near the branch sealing end. A heating pipe and a glass pipette tube in which an ultrasonic transmitter for measuring the hardness of the sucked egg cell surface and / or an ultrasonic vibrator for internal stirring are arranged at the straight end, and the heating section is heated. A power supply unit is provided, and the heating unit is energized and heated in advance for a predetermined period of time, thereby expanding the internal air in the glass pipette tube and generating a positive pressure at the tip. After the egg cells or microcapsules such as the above are brought into close contact with each other, the energization is cut to bring the glass pipette tube into a cooled state, the internal air is reduced, and a negative pressure is generated at the tip to generate the egg cells or microcapsules. Minute object suction holding device which is characterized in that so as to suck and hold the cell.

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