JP3135969B2 - Particle launcher - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a particle emitting device, and more particularly, to a particle emitting device used for introducing a gene fragment into plants, animals, microorganisms and the like.

[0002]

2. Description of the Related Art In recent years, research on genes has become active,
Many studies have been made on introducing gene fragments into plants, animals, microorganisms, and the like. Conventionally, the introduction of this gene fragment has been performed by mating, injection, electrical stimulation, drug treatment, or the like. Or it has recently, has been a method of introducing a gene fragment itself to biological cells of interest directly, is the prototype so-called particle gun method is by researchers of the United States of Cornell University development.

[0003] One of the methods is to attach a gene fragment of interest to fine particles such as tungsten or gold, accelerate it with the force of explosives, penetrate the cell membrane of the cell wall of a plant cell, and directly transfer the gene fragment into cells. As shown in FIG. 9, a synthetic ammunition created by inserting a plastic projectile 82 such as nylon or polyethylene into an empty gun 81 of explosives is used. A fixed amount of a suspension of DNA and a salt solution is placed, loaded into a shotgun 83, and driven toward cells and tissues. The sample cells, tissue 84, are located below the muzzle of the gun,
At the end of the muzzle is a stopping plate 85 having a small-diameter hole. The projected projectile 82 itself hits the stopping plate 85 and stops, but droplets containing plasmid DNA, tungsten fine particles and the like are radially fired by inertia, hit sample cells and the like, penetrate the cell wall, and enter the cells. Is done. Reference numeral 86 denotes an air vent for facilitating the movement of the projector 82. Also, the portion from the muzzle to the sample cells is housed in a closed container so that the entire device can be fired under reduced pressure.

As another method for directly implanting gene fragments into cells, a method using an air gun type apparatus as shown in FIG. 10 is also known. This method is based on the application of an air rifle firing mechanism. It has been reported that the maximum acceleration pressure of the projectile is about 240 kg / cm ² , and the initial velocity is about 1.3 times the sound velocity. . In this apparatus, first, a projectile 92 on which gold particles 91 coated with DNA are placed is inserted from the side of a stopper 93 and pushed down to the lowermost part of a barrel 94. Thereafter, the plunger pump 95 is pulled back backward by a traction mechanism (not shown), and air is introduced into the plunger 96. By triggering a trigger (not shown), the plunger pump 95 rapidly advances in the direction of the barrel 94 and moves to the air chamber 97.
Accumulates high-pressure compressed air inside. By projecting the compressed air in the air chamber 97 by the trigger 98, the projectile 92 is fired upward, and hereinafter, similarly to the shotgun type described above, the projectile 9 is projected.
2 collides with the stopper 93 and deposits the gold particles 91 on the sample cells 99.
And the gold particles penetrate the cells.

[0005]

The above-mentioned fine particle emitting apparatus for introducing a gene fragment is applicable to all organisms such as plants, animals and microorganisms, and is applicable to gene expression or traits. It is very effective and widely used in research such as conversion. In particular, transformation of monocotyledonous plants, into which gene fragments were difficult to be introduced by conventional techniques, or wild plants, to which not only the introduction of gene fragments but also the basic technology and scientific knowledge for this purpose, does not exist. It is possible to open the way, and the technical application potential of this type of particle emitting device is extremely high. For example, monocotyledonous plants have a lot of important cereals, such as wheat, which is a typical example.
Very few plasmids have been established and their regeneration system has been established without the cell wall, and future research is expected.

[0006] However, the conventionally known particle emitting devices still have some points to be improved. That is, in the case of using explosives as shown in FIG. 8, since the smoke generated by explosion of explosives is generated and remains in the container each time it is used, the container and the firing device are frequently cleaned. Must be conducted, which may hinder the smooth execution of research.In addition, since explosives are used, each member must be made robust to improve the safety of the device itself. Since high accuracy is also required for the working accuracy of the barrel, it is difficult to manufacture, and inevitably an increase in the size and cost of the device is inevitable. Further, it is practically impossible to adjust the acceleration pressure for the projectile, that is, the moving member moving in the barrel, and it is extremely difficult to change the shape or size of the barrel or the moving member. The types of target cells and the like have to be naturally restricted, and the range of research has been limited.

In addition, the handling and storage of explosives generally requires careful care, which has also created one inconvenience. The air gun type shown in FIG. 9 has been released from the use of explosives, and the work of removing smoke or safety has been greatly improved. However, the means for obtaining compressed air is a so-called air gun type means. For this reason, a large moving distance of the plunger pump is required, and it is inevitable that the whole apparatus becomes large. Further, since considerable high pressure air is used as a firing source, it is necessary to integrate the barrel portion and the plunger portion by means such as integral molding or welding, and change the form or size of the barrel or the moving member. It is equally difficult,
The target was also restricted. Further, it is necessary to place the cells serving as the sample upside down in the decompression vessel, which has made it difficult to handle the sample.

[0008] Also, there has been reported a device that uses a nitrogen cylinder pressure and opens and closes the valve in a pulsed manner using an electromagnetic valve connected to a pulse generator, thereby firing a projectile. Since the entire launching device, in which members such as the projectile mounting section are configured as a single structural body, is installed and driven in one container, the entire launching device is taken out of the container when the projectile is loaded. Handling is difficult because of the necessity or considerable changes to the launcher itself when trying to drive different types of cells under different conditions. At the same time, like other launching devices, the types of cells to be applied have to be limited, and the scope of research has been limited.

The present invention eliminates the disadvantages of the above-described conventional particle emitting device, and is not only compact and easy to handle, but also applicable to many types of cells such as cells. It is an object of the present invention to provide a particle emitting device that can easily introduce a gene fragment into cells or tissues of various kinds of plants, microorganisms, animals, and the like.

[0010]

Means for Solving the Problems In order to solve the above-mentioned problems and attain the object, the present inventors have carried out many experimental studies on a fine particle emitting device. This makes it possible to obtain a sufficient firing acceleration with a lower firing pressure by setting the friction coefficient between the moving member to which the fine particles having the gene fragments are attached and the cylinder member as the barrel to a smaller value. It is perceived that when stopped by hitting the stop member at the tip of the cylinder member, the fine particles inertially move from the moving member toward the sample cell or the like with a sufficient speed and are introduced into the cell through the cell wall. Then, based on this, the present invention was completed.

That is, the present invention provides a perforated thin tubular cylinder member, a moving member which moves inside a hole of the cylinder member capable of adhering fine particles having gene fragments and the like to an end thereof, and a chamber which can constitute a decompression chamber. And a cylinder member having one end detachably attached to a pressure fluid source via a pressure regulating valve and an opening / closing valve mechanism, and the chamber member detaching the other end of the cylinder member. It has an insertion opening that can be freely inserted and held in an airtight state, and the moving member to which fine particles and the like are attached by the energy from the pressure fluid source moves in the hole of the cylinder member toward the chamber. In addition, the present invention discloses a fine particle emitting apparatus characterized in that the fine particles and the like are fired at a lower end of a cylinder to a sample such as a cell or the like located in a chamber.

In the present invention, it is extremely possible that the moving member is held in a state in which the moving member has a partially non-contact area with respect to the inner hole penetrated by the cylinder member but does not drop by its own weight. This is a preferred embodiment, which makes it possible to fire the moving member at a lower pressure than before. For this purpose, the peripheral surface of the moving member itself may be roughened, and a thin leaf member having a rough surface such as a paper material may be attached to the peripheral wall of the moving member.

In the present invention, there is no particular limitation on the pressurized fluid serving as the emission source of the moving member, and any fluid having no toxicity to living organisms, plant cells, tissues, or the like serving as samples can be used. Can be used.

[0014]

In the present invention having the above-described structure, the moving member holding the gene fragment to be introduced and the cylinder member, which is the barrel, are mutually held under a low coefficient of friction. By using a relatively low pressure gas as the firing source, a sufficient introduction effect can be obtained.
Therefore, the device itself can be manufactured so as to have strength enough to withstand the pressure, and the device itself can be reduced in size in addition to having a simple structure as a whole, and the pressure can be easily controlled. be able to. In addition, as a result, the chamber body for accommodating the cells and the like as the sample and the cylinder member for holding the gene fragments to be implanted can be configured to be easily detachable, depending on the type of the target sample. Therefore, it is possible to replace the cylinder member or the chamber member as appropriate and work by combining the most suitable ones, and it is possible to greatly expand the target range.

[0015]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in more detail based on the description of an embodiment with reference to the accompanying drawings. In FIG.
A is a chamber member, which has a container body 1 composed of a side wall 2 made of stainless steel or the like, a bottom portion 3 and a transparent acrylic resin upper lid 4, and the bottom portion 2 of the container body 1 is appropriately raised and lowered. The mechanism 5 is mounted, and the sample mounting table 6 is located on the elevating mechanism 5.

An exhaust port 7 for reducing the pressure inside the container is formed in the side wall 2 of the container body 1, and the exhaust port 7 is connected to a vacuum pump 9 via an appropriate pipe 8. Piping 8
Is provided with a solenoid valve 10, and a branch pipe 11 is erected downstream of the solenoid valve 10. A pressure relief valve 12 is provided at the tip of the branch pipe 11. The side wall 2 is also provided with an internal pressure detection port 13, which is connected to a quartz vacuum gauge 14 located outside the apparatus. The electromagnetic valve 10 is controlled based on the information of the quartz vacuum gauge 13, and the inside of the container 1 is maintained at a preset reduced pressure environment.

The side wall 2 further has an opening 16 connected to an on-off valve 15 for breaking the vacuum in the container 1 in an emergency, and a port for inserting an appropriate measuring means such as a thermocouple thermometer if necessary. 17 are formed. The port 17 is hermetically sealed when not in use. In addition, a viewing window 18 in which a transparent resin such as an acrylic resin is hermetically mounted is provided on the side wall 2.

FIG. 2 is an enlarged sectional view of the upper cover 4 of the container body 1. An opening 19 is formed in a central portion of the upper lid 4, and a part of a holding means B for forming an insertion opening for holding a cylinder member 30 to be described later in a detachable and sealed state is inserted through the opening 19. ing. The holding means B
Has a cylindrical member 21 formed with a flange 22 at an appropriate place, and is fixed to the container body 1 by fixing the flange 22 to the upper lid 4 via an appropriate fixing means. The fixing means of the upper lid 4 and the flange 22 may be a permanent fixing means such as welding or a detachable fixing means such as a screw provided that the two are fixed in an airtight state.

An upper end portion of the cylindrical member 21 is an enlarged diameter portion. The enlarged diameter portion has a thread 23 engraved on an outer peripheral surface thereof, and has an O-shaped ring 24 having elasticity and a ring press-fit inside. A cylindrical space 26 for accommodating the piece 25 is formed. The lower end of the cylindrical space 26 has a slope inclined downward in the axial direction of the cylindrical member 21 as shown in the figure.
The lower end of the ring press-in piece 25 is a slope inclined upward in the axial direction. An opening 27 through which the cylinder member 30 can be inserted is formed at the center from the position above the enlarged diameter portion of the cylindrical member 21, and a screw that meshes with a thread 23 formed on the outer peripheral surface of the enlarged diameter portion on the inner surface. Tightening member 2 with engraved shape
8 is configured to be detachably mounted.

Therefore, after the cylinder member 30 is inserted into the holding means B, the tightening member 28 is screwed into
The ring push-in piece 25 is pressed by the tightening member 28 and descends, whereby the elastic O-ring 24 is pressed toward the surface of the cylinder member 30. Due to the pressing force, the cylinder member 30 is held in a sealed state with respect to the holding means B, that is, the container body 1.

By loosening the tightening member 28, the O-shaped ring 24 is restored to its original shape by its own elastic force, and the resistance between the O-ring 24 and the cylinder member 30 is reduced. Thereby, the cylinder member 30 can freely move with respect to the holding means B. 29 is an O-ring,
It is provided as needed when more airtightness between the holding means B and the cylinder member 30 is required.

FIG. 3 is a sectional view of the cylinder member 30. The cylinder member 30 functions as a barrel and is made of engineering plastic such as stainless steel or polycarbonate, and is basically composed of a main body cylindrical portion 31, a stopper member 32 attached to the lower end thereof, and an upper end thereof. Mounting member 33 to be mounted on
Consists of The main body cylindrical member 31 includes two cylinders 34,
Although 35 is shown as being integrally connected by welding, it may be formed of a single member, or may be formed by connecting a plurality of members by screw connection. In any case, a through-hole 36 having a diameter and a surface finishing surface capable of holding a moving member holding fine particles having a gene fragment described later without free fall is formed in the inside thereof in all cases. Further, near the lower end position, an air vent 37 is formed similarly to the barrel of the conventional launching device as shown in FIG. Screws 38 and 39 are formed on the lower end and the upper end.

The stopper member 32 has an inner surface formed with a thread engaging with a thread 38 formed at a lower end of the main body cylindrical portion 31, and has a central portion formed with the main body cylindrical portion 31. A through hole 40 having a diameter smaller than the diameter of the through hole 36 is formed. In the illustrated example, the through hole 40 is formed in an inverted conical shape having a smaller diameter toward the lower side.

The attachment member 33 has a thread 39 formed on the inner surface of the lower end thereof and formed on the upper end of the cylindrical portion 31 of the main body.
A screw thread for engaging with an opening / closing valve mechanism C described later is carved at the upper end. Further, a through hole 42 is formed at the center thereof. Therefore, the main body cylindrical portion 31, the stopper member 32 mounted on the lower end thereof, and the mounting member 33 mounted on the upper end are integrally formed by screwing each other into a cylinder member having a through hole therein. And can be easily separated individually by releasing the screw engagement.

As shown in FIG. 1, the upper end of the mounting member 33 of the cylinder member 30 is connected to a conventionally used appropriate opening / closing valve mechanism C such as a solenoid valve. It is connected to a pressure fluid source E via a pressure regulating valve D commonly used in fluid control mechanisms. In FIG. 1, F
Is a stopper member for facilitating the positioning of the cylinder member 30 with respect to the container body 1, and the positioning is performed by bringing the lower end of the on-off valve mechanism C into contact with the stopper member F.

FIG. 4 shows the moving member 50. The moving member F50 is made of a member having appropriate mechanical strength and elasticity, such as Teflon, polycarbonate, polyacetal, or a specific ceramic, as shown in the drawing.
It comprises a cylindrical portion 51 and a conical portion 52 at the tip thereof. The tip 53 of the conical portion 52 becomes flat, and fine particles such as dangsten particles and gold particles having a gene fragment adhere to the conical portion 52 together with a suspension of the salt solution and the like depending on the content of the test, as in the conventional projectile. Is done. The surface of the cylindrical portion 51 has a slightly rough surface finish, that is, a rough surface. The outer diameter of the cylindrical portion 51 is partially different from that of the cylindrical portion 51 when the cylindrical portion 51 is inserted into the through-hole 36 formed in the cylinder member 30. It has an outer diameter that has a non-contact area, but is held in a state where it does not drop due to its own weight. Although not specifically shown, the rough surface of the outer peripheral surface of the moving member 50 may be formed by appropriate machining or chemical treatment of the moving member 50 itself as described above, or the outer peripheral surface of the cylindrical portion 51 may be formed. It may be formed by sticking paper, resin material, or a composite material thereof having a required surface roughness on the surface.

Next, a method of using the particle emitting apparatus according to the present invention will be described. From the state shown in FIG. 1, the tightening member 28 of the holding means B is loosened, and the frictional engagement between the insertion opening of the holding member B and the cylinder member 30 is released, so that the cylinder member 30 is separated from the chamber member A. If necessary, the cylinder member 30 and the opening / closing valve mechanism C are disengaged from each other by screwing and separated from each other, and sterilized using an autoclave or the like.
0 is horizontally mounted on an appropriate mounting table. Next, the cylinder member 3
Then, the stopper member 32 is removed from 0.

On the other hand, a fine particle having a gene fragment or a suspension thereof with a salt solution is adhered to a flat portion 53 at the tip of a conical portion 52 of a moving member 50 which functions as a bullet. Then, the moving member 50 to which the fine particles are attached
Is pressed into the through hole 36 of the cylinder member 30 to the tip of the other end by appropriate means with the fine particle attachment surface facing rearward, loaded, and then the stopper member 32 is screwed into the lower end of the cylinder member 30 again. And fix it.

On the other hand, the upper cover 4 of the chamber member A is detached from the container main body 1, and a sample into which a gene fragment is to be implanted is mounted on the sample mounting table 6 in the container main body. After that, the cylinder member 30 is reinserted into the insertion opening and positioned with respect to the sample, and then the tightening member 28 is tightened. Thereby, the cylinder member 30 is fixed to the chamber A while maintaining the airtight state.

Next, the opening / closing valve mechanism C is mounted on the upper end of the cylinder member 30 via the mounting member 33, and the vacuum pump is operated via an appropriate control mechanism.
The inside of the container body is set to a predetermined reduced pressure state, and the state is maintained as necessary. After appropriately adjusting the opening / closing time of the pressure regulating valve D and the opening / closing valve mechanism C according to the conditions of the gene fragment to be shot or the cells to be shot, the opening / closing valve mechanism C
The movable member 50 descends in the through hole 36 of the cylinder member 30 by contacting the stopper member 32 and stops. As a result of the reaction, the fine particles adhering to the tip of the moving member 50 move inertially toward the sample in the container body, penetrate the cell wall, and are introduced into the cells as in the conventional fine particle emitting device.

In the present invention, the moving member 50 and the inner wall of the through hole 36 of the cylinder member 30 are held in such a state that they have a partially non-contact area with each other but do not fall by their own weight. I have. As a result, the pressure fluid supplied from the pressure source applies a firing acceleration to the moving member and penetrates into a part of the non-contact surface between the moving members, and functions as a lubricant between the moving member and the through hole. . Thereby, in the launching apparatus of the present invention, the conventional 100 kg / cm ²
Compared to those requiring the above acceleration pressure, 20
Even when the moving member is fired at a relatively low pressure of about 50 kg / cm ^2, a sufficient acceleration can be obtained.

Since a fluid having a relatively low pressure is used, the connection between the pressure source and the cylinder member, the configuration of the cylinder member itself, the configuration of the connection between the cylinder member and the chamber member, and the like are relatively simple. As a result, as shown in the above embodiment, the cylinder member can be detachably mounted on the opening / closing valve mechanism by a simple mechanism such as a screwing mechanism. In addition, it is possible to mount the device through an insertion opening that can be detachably inserted into the chamber member and held in an airtight state. As a result, various combinations of the pressure source, the cylinder member, and the chamber member can be easily exchanged and fired at a sample such as a cell located in a chamber such as a fine particle, thereby greatly expanding a test object. It is possible to do.

[0033]

[Practical example] An example in which a gene fragment is actually introduced using the fine particle emitting device according to the present invention will be described below.

[0034]

[Practical Example 1] A young plant of tobacco ( Nicotiana tabacum ) aseptically grown in a culture apparatus (10 cm in height and 10 cm in diameter) constituting the chamber of the present invention was removed from the container. An experiment was conducted in which a gene was directly introduced into a living tobacco seedling itself by using the apparatus of the present invention while being placed on a sample stand.

A plasmid DN having a β-glucuronidase (GUS) gene in gold particles having a diameter of about 0.8 μm.
A, pBI221 (manufactured by Clonetech, USA) was adhered, and the ethanol suspension was dropped on the tip of a plastic bullet (Teflon or the like; 5φ) and dried. In this case, the weight of the plasmid DNA and the gold particles is 200 ng D
NA / 50 μg The ratio of gold particles.

Next, the distance between the tip of the cylinder (barrel) and the tobacco leaf was set to 5 cm, the acceleration pressure of the bullet was set to 24 kg / cm ^2, and gold particles were shot under reduced pressure of 100 mmHg. After transfection, the cells were cultured in the light for 24 hours, and then the GUS substrate X-Gluc (5-bromo-4-chloro-3indolyl-glucoronid
The tobacco leaves were treated with a buffer solution containing e), decolorized with 70% ethanol, and observed.

As is apparent from FIG. 7, a number of blue spots composed of cells transiently expressing the GUS gene were observed. On the other hand, no blue spot was observed from the leaves which were shot with gold particles to which no plasmid DNA was attached. The number of blue spots, which indicate the number of cells expressed after introducing the gene, which is an index of the performance of this apparatus, was about 7,000 / cm ² .

[0038]

[Indeed Example 2] Under the experimental conditions as in Example 1, the conventional most gene transfer has not been attempted rugosa (Rosa Rugosa)
An experiment for introducing a GUS gene into leaves was performed using this apparatus. As a result, as apparent from FIG. 8, many blue spots were observed, and the expression of the GUS gene was confirmed.
The number of blue spots at this time was about 5000 / cm ² .

[0039]

[Example 3] Under the same experimental conditions as in Example 1, the yacon ( Po
( G. lymnia sonchifolia ). As a result, the expression of the GUS gene was confirmed, and the number of blue spots was approximately 5000 / cm ² . It was confirmed that artificial gene transfer into Yacon was successfully performed by using this device.

[0040]

[Practical Example 4] Under the same experimental conditions as in Example 1, an experiment of introducing a GUS gene into leaves of iris ( Iris species) was performed. As a result, the expression of the GUS gene was confirmed, and the number of blue spots was 3000 to 4000 / cm ² . It was confirmed that the gene was successfully introduced into the iris of a monocotyledon by using this apparatus.

[0041]

Embodiment 5 Under the same experimental conditions as in Embodiment 1, Avena ( Av
ena sativa ), the GUS gene was introduced into leaves and embryos, and its expression was confirmed. By using this device, the gene was successfully introduced and expressed in one of the world's first important cereals, wheat.

[0042]

Description of Modifications The description of the fine particle emitting apparatus according to the present invention described above is merely an explanation of one embodiment, and there are many other modifications. For example, the connection mode between the cylinder member and the pressure fluid source is not limited to that shown in the drawing, and the pressure fluid of the pressure fluid source is supplied to the moving member loaded in the cylinder member at a predetermined pressure for a predetermined time. As long as the connection is in such a manner, all conventionally known pressure regulating mechanisms and on-off valve mechanisms can be used.

Further, the structure for detachably fixing the cylinder member to the chamber member in an airtight state is not limited to the structure using an O-ring having elasticity as shown in FIG. The mechanical coupling provided may be used, or the upper cover 4 and the holding means B may be integrally formed. Further, as for the cylinder member, as shown in FIG.
The upper end of a may be configured to be flat, and the mounting member 33 may be mounted on the flat surface via an O-ring 60 by detachable fixing means (not shown). An O-ring 61 may be attached to a portion to be supported for the purpose of securing the support and stability of the moving member 50. In this case, it is easy to load the moving member from the upper end of the cylinder member 30 and the operability is further improved (even with the cylinder member shown in FIG. Of course, it is possible to remove the 41 and load the moving member from above, as well as to position the O-ring).

The chamber members are also shown in the drawing.
Although only one insertion opening for mounting the cylinder member is shown in the upper lid portion, it will be easily understood that a similar insertion opening can be provided in the side wall portion and also in the bottom portion in some cases. For example, if a similar insertion opening is provided at the bottom and a cylinder member is attached to it, it will be possible to introduce gene fragments into the back as well as the front of tobacco leaves, and to conduct more thorough tests Can be.

Further, in the particle emitting apparatus of the present invention, since the chamber member and the cylinder member can be easily attached and detached, the chamber member is formed as a bottomless chamber Aa as shown in FIG. After the bottom portion of the bottomless chamber member is brought into close contact with the animal or plant itself, the gene fragment can be introduced by inserting and fixing the cylinder member and reducing the pressure inside. In FIG. 6, reference numeral 7a denotes a degassing port, and 62 denotes a flexible member for further ensuring the confidentiality between the target animal and plant and the chamber member.

[Brief description of the drawings]

FIG. 1 is a partial cross-sectional view showing a configuration of a main part of an embodiment of a particle emitting device according to the present invention.

FIG. 2 is an enlarged sectional view showing a cylinder member insertion opening.

FIG. 3 is an enlarged sectional view of a cylinder member.

FIG. 4 is a view showing a moving member.

FIG. 5 is a partially enlarged sectional view showing another embodiment of the cylinder member.

FIG. 6 is a perspective view showing another embodiment of the chamber member.

FIG. 7 is a view showing the expression of a gene introduced into tobacco leaves (photograph showing the form of an organism).

FIG. 8 is a view showing the expression of a gene introduced into the leaves of Hamanas (a photograph showing the form of an organism).

FIG. 9 is an explanatory view showing a conventional particle emitting device.

FIG. 10 is an explanatory view showing another conventional particle emitting device.

[Explanation of symbols]

A: chamber member, 1: container body, 6: sample table, 9
... Vacuum pump, 30 ... Cylinder member, C ... On-off valve mechanism,
D: pressure regulating valve, E: pressure fluid source

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-3-251187 (JP, A) JP-A-1-90393 (JP, U) WO 91/18991 (WO, A1) (58) Fields surveyed (Int.Cl. ⁷ , DB name) C12M 1/00-1/34

Claims (3)

(57) [Claims] 1. A cylindrical member having a perforated tube, a moving member which moves inside a hole of the cylinder member to which fine particles having gene fragments can be attached to one end thereof, and a chamber member which can constitute a decompression chamber. The moving member is located in the inner hole of the cylinder member.
On the other hand, in a state where there is a partially non-contact area with each other,
But can be held without falling by its own weight.
And one end of the cylinder member is detachably attached to a pressure fluid source via a pressure regulating valve and an opening / closing valve mechanism, and the chamber member vertically moves the cylinder member.
Inserted upright posture the other end freely leave at To and has an insertion opening which can be held in an airtight state, the moving member attached to fine particles by the energy from the pressure fluid source the cylinder A fine particle launching apparatus which moves inside a hole of a member toward the chamber, and discharges the fine particles and the like to a sample such as a cell located in the chamber at a lower end of a cylinder. 2. The moving member according to claim 1, wherein a thin leaf member having a rough surface is attached to a peripheral wall of the moving member.
The particle emitting device according to claim 1. 3. The particle emitting device according to claim 2, wherein said thin leaf member is a paper material.