WO1992013062A1

WO1992013062A1 - Jetting device for treating cell

Info

Publication number: WO1992013062A1
Application number: PCT/JP1992/000028
Authority: WO
Inventors: Minoru Igari; Minoru Takahashi; Ichiro Takakura; Naohiko Takahashi; Masahiko Kitayama
Original assignee: Nippon Zeon Co., Ltd.
Priority date: 1991-01-18
Filing date: 1992-01-16
Publication date: 1992-08-06
Also published as: AU1168792A

Abstract

A gas storing room (21) and a resisting piece (32) are provided between a gas generating part (20) and a cylinder (22) within which a piston (23) moves. The resisting piece (32) comprises a board made of paper to be broken when subjected to the gas pressure and a partitioning plate having an opening. Such a structure as above adapts the piston (23) to move forward not immediately after generation of gas but after the rise of gas pressure in the gas storing room (21) and thus the piston is accelerated to high speed by the full use of the high pressure, whereby jetting speed of a cell treating material (24) is increased. Another structure to arrange a tank (89) adjustable in volume between the gas generating part (20) and piston (23) is also possible. In this structure, gas from the gas generating part (20) enters the tank (89) and therefore the pressure in the rear of the piston (23) is regulated according to the volume of the tank (89) so that the speed of the piston, that is, the jetting speed of the cell treating material (24) may be regulated.

Description

Akira glue Cell processing injection equipment Technical field

The present invention relates to a cell processing injection device used to cause transformation of cells.

That is, this injection device, for example, injects a cell treatment material such as metal fine powder to which genetic material is attached at a high speed, and injects the genetic material into cells by applying the sample to cells, including cells. It is used for high-speed injection of cells to cause cell fusion. Background art

As an injection device for cell treatment, plastic piston with cell treatment material attached to its tip is accelerated at high speed using explosives, compressed air, etc., and collides with a plate with small holes, etc. There is known one in which a processing material is injected.

With this conventional apparatus, it is difficult to increase the speed of the piston, that is, the cell treatment material. Experiments have shown that, even if the amount of explosive used is greatly increased, the speed of the device using explosives is negligible. This is due to the fact that the biston moves with little resistance to the cylinder, causing it to travel forward and collide before the gunpowder burns enough to generate high pressure. Because That is, production and handling For the sake of simplicity, the structure is such that very little frictional force is generated between the piston and the cylinder. As a result, the piston cannot stay at the equipment position until the explosive combustion speed reaches its maximum and the gas pressure rise rate at the rear of the piston reaches a peak, and the explosive combustion occurs. In the very early stage, the gas pressure at the rear of the piston cannot rise sufficiently because the piston moves forward in the barrel and is discharged outside the barrel. Under these conditions, the amount of explosive charge has little effect on piston speed.

Similarly, when using compressed air, if the injection valve is opened, the biston will advance before the pressure behind it becomes sufficiently high, making it difficult to increase the speed. is there.

In this apparatus, the means for adjusting the speed of the piston, that is, the cell treatment material, is not particularly effective in an apparatus using explosives except for changing the amount of explosive. There is only a method of adjusting the gas pressure in an apparatus that uses a gas. Therefore, there is a drawback that speed adjustment is difficult and the adjustment range is narrow. Disclosure of the invention

Therefore, the present invention provides a cell processing injection device having a new configuration capable of sufficiently increasing the speed of the piston, that is, the cell processing material, and according to the piston speed. It is an object of the present invention to provide a cell processing spray device capable of easily and widely adjusting the spray speed of a cell processing material. O

The cell processing injection device of the present invention includes a gas generator that generates a high-pressure gas instantaneously, a gas reservoir that is connected to the gas generator, and that receives the gas, and a gas reservoir that is connected to the gas reservoir. It has the same or smaller cross-sectional area as the gas reservoir, stores the piston with the cell treatment material attached to the tip, and moves the piston toward the tip by the gas from the gas reservoir. The cylinder has a small hole and a small hole, and is provided on the tip of the cylinder or on the extension of the small hole. The cylinder collides with the piston to eject the cell treatment material from the small hole to the sample. And a resistor that is provided between the gas reservoir and the cylinder, and that adds resistance to the flow of gas to the cylinder. The device can have a higher piston speed than the conventional one.

Further, in the injection device for cell treatment of the present invention, the volume can be adjusted between the gas generating section and the piston in the cylinder, and the flow of the gas to the cylinder can be reduced. It may be provided with a tank for receiving and storing parts. In this case, the piston speed can be arbitrarily adjusted by adjusting the tank volume of the cell processing injection device.

The gas generating section of the cell processing injection device may be composed of an explosive and an ignition mechanism for igniting the explosive or a high-pressure gas tank and an injection valve connected thereto. it can.

The resistor is provided to partition the gas flow, A plate made of paper, plastic, or the like that is broken by gas can be used, and the shape can be a flat plate, a conical shape projecting toward the gas generating portion, or the like. In addition, a partition plate provided so as to partition the gas flow and having an opening having a cross-sectional area smaller than the cross-sectional area of the cylinder may be used. The opening may be formed of one hole, or may be cup-shaped, and the opening may be formed of a plurality of holes.

Further, the injection device for cell treatment of the present invention further comprises an outlet hole for allowing a part of the gas flow to the cylinder to flow between the gas generating portion and the piston in the cylinder; The injection speed may be adjustable by providing a pressure regulating valve connected to this outlet hole to open and release gas when the pressure reaches a preset value. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a perspective view of a first embodiment of the present invention,

Fig. 2 is a longitudinal sectional view of the main part of the example,

FIG. 3 is a longitudinal sectional view of a main part of the second embodiment,

FIG. 4 is a longitudinal sectional view of a main part of the third embodiment,

FIG. 5 is a longitudinal sectional view of a main part of the fourth embodiment,

FIG. 6 is a longitudinal sectional view of a main part of the fifth embodiment,

FIG. 7 is a longitudinal sectional view of a main part of the sixth embodiment.

Hereinafter, embodiments of the present invention will be specifically described with reference to the drawings. explain.

1 and 2 show an injection device 10 for cell treatment according to a first embodiment of the present invention. This device 10 is entirely covered with a detachable case 12 mounted on a base 11. By suctioning the internal air from the suction port 13 with the case 12 covered by a vacuum pump (not shown), the entire interior of the case 12 can be reduced in pressure.

A support 14 is provided on the base 11, and a holder 15 is mounted on the support 11 so that a vertical position can be adjusted by operating a holding screw 16.

The holder 15 supports the cylinder 17, and an empty gun cartridge 18 containing gunpowder can be loaded into the upper end of the cylinder 17. Is equipped with an empty gun cartridge 18 and is provided with an ignition mechanism 19 for igniting, and the empty gun cartridge 18 and the ignition mechanism constitute a gas generating section 20. . The ignition of the ignition mechanism 19 is operated by a solenoid, and the operation is performed outside the case 12.

The inside of the cylinder 17 is a gas reservoir 21, and a cylinder 22 is detachably screwed to a tip of the gas reservoir 21. The cylinder 22 houses the piston 23 inside. The piston 23 is made of plastic, and a rib can be protruded around the rear to press the inner surface of the cylinder 22 to hold the position, and a recess is formed at the tip. Here, the cell treatment material 24, for example, tungsten fine powder with genetic material attached thereto, for example, It is attached by the surface tension of water.

A root of a front cover 25 is detachably attached to a tip end, that is, a lower end of the cylinder 22 by a bayonet-type screw. The front cover 25 covers the sample 26 placed on the base 11 and holds the injection section 27 between the front end of the cylinder 22 and the injection section 27. The jetting portion 27 has a small hole 28, and collides the piston 23 to eject the cell treatment material 24 from the small hole 28.

Exhaust holes 29, 30, ... are provided near the tip of the cylinder 22, and exhaust holes 31, ... communicating with the exhaust holes 30, ... are also provided at the base of the front cover 25. ... are provided. A resistor 32 is provided between the tip of the cylindrical body 17 and the cylinder 22 so as to partition the gas flow between the gas reservoir 21 and the cylinder 22. I have. The resistor 32 is a flat plate made of a plate made of paper, plastic, or the like that can be broken by gas.

Next, the operation of the cell processing injection device having such a configuration will be described.

First, prepare as follows. Place sample 26 on base 1 1. The piston 23 with the cell treatment material 24 attached to the cylinder 22 is housed, and attached to the body 17 with the resistor 32 interposed therebetween. Load empty cartridge 18 and secure holder 15 at the appropriate height. Cover Case 12 and reduce the pressure inside Case 12.

Next, the ignition mechanism 19 is operated. When you do this, the empty cannon High-pressure gas is generated from the gunpowder in the cartridge 18, and this gas fills the gas reservoir 21, destroys the resistor 32, proceeds into the cylinder 22, and removes the screw. The ton 23 is pressed to accelerate at a high speed and collide with the injection part 27. Due to this collision, the cell treatment material 24 is ejected through the small hole 28, and the cell treatment such as injecting the genetic material into the cells is achieved by hitting the sample 26. At this time, the air remaining in front of the piston 23 is exhausted from the exhaust holes 29,…, 30,, 31,…. Once this device is used, the piston 23 and resistor 32 are damaged and must be replaced with the empty gun-to-trigger 18.

FIG. 3 shows an injection device 40 for cell treatment according to a second embodiment of the present invention. The other configuration is the same as that of the device of the first embodiment except that the antibody 32 of the cell processing injection device 10 of the first embodiment is replaced by another resistor 41. The same reference numerals are given to the corresponding parts of the device of the first embodiment. The resistor 41 of this device is made of a plate made of paper, plastic, or the like, which bursts by gas, and has a circular shape protruding toward the gas generating portion. ing.

The cell processing injection device 40 of the second embodiment is also used and operates in the same manner as the device 10 of the first embodiment. Since the resistor 41 of this device has a conical shape protruding in the gas generation direction, it can be prevented from breaking down to a pressure higher than that of the resistor 32 in the first embodiment. At higher pressures, flow gas through pistons 23 and It is possible to accelerate the speed of the operation.

FIG. 4 shows a cell processing injection device 50 according to a third embodiment of the present invention. The other configuration is the same as that of the device of the first embodiment, except that the antibody 32 of the injection device for cell treatment 10 of the first embodiment is replaced by another resistor 51. The same reference numerals are given to the corresponding parts of the device of the first embodiment. The resistor 51 of this device is formed of a flat partition plate for partitioning the gas flow, and has an opening 52 formed of the first hole. The cross-sectional area of the opening 52 is smaller than the cross-sectional area of the cylinder 22.

This device 50 is used in the same manner as the device 10 of the first embodiment, and operates almost in the same manner. The resistor 51 of this device does not break, but gives resistance to the gas flow through the opening 52. Since the gas generation in the gas generation section 20 is extremely fast, high-pressure gas can be stored in the gas storage chamber 21 only by the flow resistance of the opening 52. Therefore, after the gas once becomes high pressure, it flows into the cylinder 22 and the piston 23 advances, so that the piston 23 is accelerated at a high speed.

According to experiments, the collision velocity of the piston was 660 mZs when no resistor was used, whereas this resistor 51 (the cross-sectional area of the opening was the cross-sectional area of the cylinder) The time was 1800 ms when using (half).

FIG. 5 shows an injection device 60 for cell treatment according to a fourth embodiment of the present invention. The cylinder of the injection device 10 for cell treatment of the first embodiment In this example, the body 17 is replaced with a cylindrical body 61 having a large diameter at the base, and another resistor 62 is used in place of the resistor 32, and the other configuration is the same as that of the device of the first embodiment. Similarly, the same reference numerals are given to the corresponding parts of the device of the first embodiment. The resistor 62 of this device is formed of a cup-shaped partition plate for partitioning the gas flow, and has an opening 63 formed of a plurality of holes. The cross-sectional area is smaller than the cross-sectional area of the cylinder 22. An intermediate chamber 64 having a larger sectional area than that of the cylinder 22 is formed between the gas reservoir 21 and the cylinder 22, and the cylinder 22 is formed in the intermediate chamber. It will be connected to the gas reservoir 21 via 6 4.

This device 60 is used and operates in the same manner as the device 50 of the third embodiment.

FIG. 6 shows a cell processing injection device 70 according to a fifth embodiment of the present invention. This device 70 is capable of adjusting the injection speed of the device 50 of the third embodiment. In this device 70, the holder 15 and the cylinder 17 of the third embodiment are replaced with holders 71 and a cylinder 72 of another configuration, and the other parts are the device 50 of the third embodiment. The same components as those in the third embodiment are denoted by the same reference numerals as those in the third embodiment. The holder 71 has a small diameter at the lower end, and the cylindrical body 72 has an outflow hole 73 on the peripheral surface near the empty gun cartridge 18 and a screw groove 74 on the outer periphery. Further, in this device 70, an adjusting ring 75 is screwed into the screw groove 74 so that a spring 76 pressed by the adjusting ring 75 can move up and down on the outer periphery of the cylindrical body 72. Mating and outlet holes 7 3, A pressure regulating valve 78 is added to press the movable ring 77 having a substantially force-up shape which receives gas from… against the housing 71 o

This device 70 is also used in the same manner as the device 50 of the third embodiment, and operates to eject the cell treatment material 24. In this device 70, since there is a pressure regulating valve 78, the movable ring 77 is pressed with an appropriate strength by operating the adjusting ring 75 in advance, so that the inside of the gas reservoir 21 is When the pressure becomes constant, the gas entering the movable ring 77 from the outlet holes 73, ... pushes down the movable ring 77 against the panel 76, and the gas is discharged to the outside. For this reason, the speed of the piston 23, that is, the injection speed of the cell treatment material 24 is adjusted.

FIG. 7 shows a cell processing injection device 80 according to a sixth embodiment of the present invention. This device 80 is also capable of adjusting the injection speed of the device 50 of the third embodiment. In this device 80, the cylinder 17 and the cylinder 22 of the third embodiment are replaced with a cylinder 81 and a cylinder 82 of another configuration, and the other parts are the devices of the third embodiment. The configuration is the same as that of 50, and the components are denoted by the same reference numerals as the corresponding components of the device of the third embodiment. The cylindrical body 81 has an outflow hole 83 on the peripheral surface near the empty gun cartridge I 8, and forms a peripheral groove 84 with the holder 15. A keyway 85 is provided on the peripheral surface. Further, in this device 80, an adjusting ring 86 and a movable ring 87 are added. The adjustment ring 86 has a substantially inverted cup shape, has a thread groove 88 on the inner surface, and is rotatably fitted to the peripheral groove 84. The movable ring 8 7 The cylinder 82 is fitted to the outer periphery so as to be vertically movable and screwed into a screw groove 88 to form a tank 89 with the inside of the adjusting ring 86. The movable ring 87 has a small screw 90, and the small screw 90 is fitted into the key groove 85 so that the movable ring 87 cannot be rotated. Thereafter, by rotating the adjustment ring 87, the volume of the tank 89 can be adjusted by moving the adjustment ring 87 up and down.

This device 80 is also used in the same manner as the device 50 of the third embodiment, and operates to eject the cell treatment material 24. Since the device 80 has a tank 89, the tank 89 can be adjusted by operating the adjusting ring 86 in advance to adjust the capacity of the tank 89. Receives an appropriate amount of gas from the outflow holes 83,...-The pressure in the gas reservoir 21 can be limited, so that the speed of the piston 23, that is, the injection speed of the cell treatment material 2, is adjusted. Is done.

The injection of the cell processing material 24 by the cell processing injection device 80 will be described.

When the ignition mechanism 19 is operated with the same preparation as the device 10 of the first embodiment, high-pressure gas is generated from the gunpowder in the empty gun cartridge 18, and this gas is used as a resistive element. 5 Due to the flow resistance of the opening 52, the gas reservoir 21 and the tank 89 are filled, and then the cylinder 82 is moved to the piston 23. Press to accelerate at high speed and collide with the jetting part 27. Due to this collision, the cell treatment material 24 is ejected through the small holes 28 and hits the sample 26 to inject genetic material into the cells. Cell processing is realized. At this time, the air remaining in front of the piston 23 is exhausted from the exhaust holes 29,, 30…, 31,….

That is, the tank 89 adjusts the speed of the piston 23, that is, the injection speed of the cell treatment material 24, by receiving the gas and adjusting the pressure behind the piston 23. Therefore, the injection speed decreases as the volume increases.

The resistor 51 provides resistance to the flow of gas to the piston 23, and the piston proceeds at a low speed before the gas generator 21 yet generates enough gas. This is intended to prevent this from occurring and to increase the speed of the piston and expand the speed adjustment range.

Although the embodiments of the cell processing injection device according to the present invention have been described above, the present invention may have various configurations other than these examples. For example, all of the above examples use explosives as the gas generating unit, but use high-pressure gas such as one that blows out compressed air with a blow-off valve, similar to an air gun. Things can also be used. The injection section may be provided at the tip of the cylinder as shown in the figure, or may be provided at a distance from the tip of the cylinder. If the injection speed of the resistor is to be increased and widened, the low-speed device can be omitted. Any configuration, such as an inserted one, can be used. The configuration of the pressure regulating valve and tank for adjusting the injection speed is also the same as the above example. It is not limited.

For example, the configuration of the tank may be such that a part of the cylinder is made large in diameter and the two large diameter parts are expanded and contracted. In short, it is necessary to receive gas and adjust the volume. It is optional as long as it is possible.

In particular, the positions of the outflow holes used for these may be provided not only upstream of the resistor as shown in the figure, but also downstream. In addition, the illustrated device is used under reduced pressure, but may be used under normal pressure depending on the application. The invention's effect

As described above, the injection device for cell treatment of the present invention has the resistor, so that the gas from the gas generating unit is temporarily stored in the gas storage chamber instead of immediately pressing the piston. Therefore, the conventional operation in which the piston proceeds to complete the injection before the gas becomes sufficiently high in pressure is improved, and the piston advances after the gas becomes high in pressure, and the high speed operation is performed. Injection speed. In particular, when explosives are used in the gas generating section, the combustion speed of the explosives is accelerated by the high pressure in the gas reservoir, so that the injection speed is extremely high. In addition, since the injection speed becomes high, if the speed can be adjusted using a pressure regulating valve or tank, the adjustment range will be wide and it will be used in various application fields. Become.

Claims

Contract

1. A gas generator that generates high-pressure gas instantaneously, and a gas reservoir that is connected to the gas generator and receives gas

When,

Connected to this gas reservoir, it has the same or smaller cross-sectional area as the gas reservoir, and stores a piston with the surrounding cell treatment material attached to the tip, and receives the gas from the gas reservoir. A cylinder that advances the piston toward the distal end,

An injection section having a small hole, which is provided on the tip or extension of the tip of the cylinder, and which collides with a piston to inject the cell treatment material from the small hole to the sample; A resistor provided between the cylinder and the cylinder to add resistance to the flow of gas to the cylinder;

An injection device for cell treatment, comprising:

2. The injection device for cell treatment according to claim 1, wherein the gas generator comprises an explosive and an igniter for igniting the explosive.

3. The injection device for cell treatment according to claim 1, wherein the gas generating unit comprises a high-pressure gas tank and a jet valve connected thereto.

4. The cell treatment device according to claim 1, 2 or 3, wherein the resistor is provided to partition a gas flow, and is made of a plate made of paper, plastic, or the like, which is broken by the gas. Injection device.

5. The spray device for cell treatment according to claim 4, wherein the plate is a flat plate.

6. A plate-shaped body protruding toward the gas generator The injection device for cell treatment according to claim 4, which is:

7. The cell according to claim 1, 2 or 3, wherein the resistor comprises a partition plate provided to partition a gas flow and having an opening having a cross-sectional area smaller than that of the cylinder. Injection for processing

8. The injection device for cell treatment according to claim 7, wherein the partition plate has a flat plate shape, and the opening is formed by one hole.

9. The injection device for cell treatment according to claim 7, wherein the partition plate is cup-shaped, and the opening has a plurality of holes.

10. An outlet provided between the gas generating section and the piston in the cylinder to allow a part of the gas flow to the cylinder to flow, and connected to the outlet, The cell processing injection device according to any one of claims 1 to 9, further comprising a pressure regulating valve that opens when the pressure reaches a preset value and releases the gas to the outside, and the injection speed can be adjusted.

1 1. A tank is provided between the gas generating section and the piston in the cylinder, the volume of which can be adjusted, and a tank that receives a part of the gas flow to the cylinder and stores it. The cell processing injection device according to any one of claims 1 to 9, wherein the injection device has an adjustable injection speed.

1 2. A gas generator that generates high-pressure gas instantaneously,

A gas storage chamber connected to the gas generating section for receiving gas;

A piston connected to this gas reservoir, having the same or smaller cross-sectional area as the gas reservoir, and having a cell treatment material attached to the tip. A cylinder for storing the piston and advancing the piston toward the distal end by gas from the gas reservoir.

An injection unit having a small hole, which is provided on the tip or extension of the tip of the cylinder, and which collides with a piston to inject the cell treatment material from the small hole into the sample;

The volume can be adjusted between the gas generator and the piston in the cylinder, and a tank that receives and stores a part of the gas flow to the cylinder.

An injection device for cell treatment, comprising: