JPH0678743A - Filter for substance introducing system and substance introducing system - Google Patents

Abstract

PURPOSE:To provide a substance introducing system designed to trap tailings and/or thready chips produced when nylon projectiles impinged on a stop plate to prevent the pollution of the area where a specimen is placed, along with protecting cells as the specimen, thereby facilitating filter maintenance. CONSTITUTION:The substance introducing system is made up of (A) a vessel 1 so designed that a vacuum chamber 10 is provided, the top face 14 is provided with an opening 140, and a specimen is placed at the lower part and the other wall surface is provided with an opening 13 communicating with a vacuum suction pump 12, (B) a mechanism 2 fixed firmly to the top face of the vessel 1 and to emit nylon projectiles, (C) a stop plate 4 located at the support 31 of a member 30 (a stop plate support) and to receive nylon projectiles emitted from the mechanism 2 and make a substance introduced fly via a hole 41 to the specimen T, and (D) a filter member 5 located in between the stop plate 4 and the specimen T and to trap tailings scattered when nylon projectiles impinged on the stop plate.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention is directed to hitting a stop plate supported by a stop plate supporting device in a vacuum chamber with a bullet piece so that a gene or other introduced substance carried on the bullet piece is a living cell or tissue. The present invention relates to a filter device for a substance introduction device and a substance introduction device to be introduced into another sample.

[0002]

2. Description of the Related Art Conventionally, as a material introduction apparatus of this type disclosed in Japanese Patent Laid-Open No. 1-227764, a vacuum chamber C formed in a housing means H has upper and lower chambers with a horizontal portion M as shown in FIG. Divided into UC and DC,
Upper and lower chambers UC and DC are surge tanks S
Shut-off valves UV, DV and filters UF, DF communicating with
Independent chambers that are independently vacuum-controlled by the conduits UL and DL having A collision receiving mechanism R is arranged at the opening of the horizontal portion M, and the multi-shot piece ejected by the propulsion means P collides with the impact stopping member R and is to be released from the central opening O into the microbial sample. In some cases, the small bullets covered with are sent to the sample biological material BS, and the multiple bullets block the central opening O and seal the upper and lower chambers UC and DC.

[0003]

In the above-mentioned conventional substance introducing apparatus, the inside of the vacuum chamber C is divided into the upper and lower independent vacuum chambers UC and DU by the horizontal portion M. Therefore, the upper and lower vacuum chambers UD and Since the DC vacuum control needs to be independently performed by the shutoff valves UV and DV and the conduits UL and DL, the vacuum control operation is difficult and the structure is complicated.

Further, in the above-mentioned conventional apparatus, since there is no filter device between the impact stop member R and the biological material BS, dust or thread-like debris generated when the small bullet collides with the impact stop member R. Has a problem that it reaches the biological material BS which is a sample and damages, sometimes destroys or kills the cells of the biological material BS which is a sample.

Further, in the above conventional apparatus, the upper and lower chambers U
By the upper and lower conduits UC communicating with C and DC, and the upper and lower filters UF and DF arranged in DC, the upper and lower chambers U are collided with the impact stopping member R of the multi-shot piece and explosive of explosives.
Since it collects the dust and the like generated in C and DC,
The maintenance of the two upper and lower filters UF and DF is difficult,
There was a problem that the structure became complicated.

Therefore, the inventors of the present invention support a stop plate for stopping the projectiles in a single vacuum chamber from which the horizontal members for partitioning the vacuum chamber in the upper and lower parts are removed by a stop plate supporting mechanism having a small area. Focusing on the technical idea of arranging the sample so as to face the stop plate and capturing the dust generated when the projectile collides with the stop plate between the stop plate and the sample to prevent it from reaching the sample. At the same time, the present invention was focused on the technical idea that thread-like fragments generated by the collision of bullets with the stop plate are collisionally locked by the locking portion. Furthermore, as a result of repeated research and development, the inventors of the present invention have confirmed that in the filter device for substance introduction device and the substance introduction device, debris and thread-like debris generated when the bullet collides with the stop plate reach the sample. A book that achieves the purpose of preventing the damage of cells of the sample, sometimes destroying or killing the sample, introducing the substance to be introduced into the sample, facilitating maintenance of the filter device and simplifying the structure. The invention was reached.

[0007]

A filter device for a substance introducing device according to the present invention (a first invention according to claim 1) is supported by a stop plate supporting device in a vacuum chamber of the substance introducing device and is ejected from a firing mechanism. It is composed of a stop plate for receiving a shot piece provided with the emitted introduction substance and flying the introduction substance toward the sample, and a filter member arranged between the sample placed in the vacuum chamber, It is configured such that dust and debris generated when the piece collides with the stop plate and prevents the piece from reaching the sample.

The filter device for a substance introducing device according to the present invention (the second invention according to claim 2) is the mesh device according to the first invention, wherein the filter member is arranged below the stop plate and above the sample. It is composed of a member, and has a locking portion having a small area in the center thereof, and is configured to collide and lock the thread-like fragments generated by the collision of the bullet with the stop plate.

The substance introducing apparatus of the present invention (the third invention according to claim 3) is a container which forms a vacuum chamber, has an opening, and in which a sample for introducing the substance is placed, and the substance introduced through the opening. The firing mechanism for firing the provided bullet pieces, the stop plate support device in which the support part is arranged at a distance from the sample fixed in the container and placed in the vacuum chamber, and the support part of the stop plate support device A stop plate and a stop plate support device, which is a member having a hole portion in the center, which receives a piece of bullet fired from a firing mechanism and at the same time causes the introduced substance to fly toward the sample through the hole portion. The filter device is composed of a filter member arranged between the sample and the filter, and filters the dust and the like generated when the bullet hits the sample plate so as not to reach the sample.

[0010]

According to the filter device for a substance introducing device of the first aspect of the present invention having the above-described structure, the projectiles having the introducing substance emitted from the firing mechanism of the substance introducing device filter the dust scattered when the projecting plate collides with the stop plate and is received. It is what the member captures.

In addition to the effect of the first aspect of the invention, the filter device for a substance introducing apparatus of the second aspect of the present invention having the above-mentioned structure has a thread-like shape which is scattered from the hole of the stop plate when the projectile collides with the impact plate. The debris is locked by colliding with the central locking portion.

In the substance introducing device according to the third aspect of the present invention having the above-mentioned structure, the projecting mechanism ejects a piece of the bullet containing the introduced substance,
The projectile collides with the stop plate arranged in the vacuum chamber and is received by the supporting portion of the stop plate supporting device.
The introduced substance is made to fly toward the sample through the hole to introduce the substance into the sample, and at the same time, a piece of dust with the introduced substance emitted from the firing mechanism collides with the stop plate and scatters when it is received. Is captured by the filter member.

[0013]

In the filter device for a substance introducing device according to the first aspect of the present invention, which has the above-described action, the filter member captures the scraps scattered when the bullet pieces collide with the stop plate and are received, so that only the substance is introduced into the sample. In addition to the effect that it is possible to prevent, it is possible to prevent the residue generated when the bullet hits the stop plate from reaching the sample, and to facilitate the maintenance of the filter device and simplify the structure. .

In addition to the effect of the first aspect of the present invention, the filter device for a substance introducing apparatus of the second aspect of the present invention that has the above-described action has the locking portion for the debris and thread-like debris generated when the projectile piece collides with the stop plate. It has an effect of preventing the cells of the sample from being locked and reaching the sample to prevent the cells of the sample from being damaged, sometimes destroyed or killed.

In the substance introducing device of the third invention having the above-mentioned action, the filter member captures the dust scattered when the projecting piece collides with the stop plate and is received, so that only the substance can be introduced into the sample. In addition, there is only one vacuum chamber in the container, the operation of vacuum control of the vacuum chamber is simple, and the structure is also simple, and the structure of the filter device can be easily maintained. It has the effect of simplifying.

The substance introducing apparatus of the third invention having the above-mentioned action exhibits the effects of the first invention, and at the same time, removes the dust passing through the hole of the stop plate, the thread-like fragments and the dust reaching in other time. Since it is captured and blocked by the filter member of the filter device, it is possible to introduce only a necessary substance into the sample.

[0017]

Embodiments of the present invention will now be specifically described with reference to the drawings.

(First Embodiment) A first embodiment relates to a filter device for a substance introducing device according to an embodiment of the first invention and a substance introducing device according to an embodiment of the third invention.
Hereinafter, this is simply referred to as a substance introduction device.

In the substance introduction device of the first embodiment, as an example, a sample T of a young leaf slice of corn, which is the second leaf after one week of germination, was loaded with a plasmid incorporating the 35S / GUS gene on tungsten particles. Shoot with something,
It is to be introduced.

As shown in FIG. 1, the substance introducing apparatus of the first embodiment forms a vacuum chamber 10, an opening 14O is formed in an upper surface portion 14, a sample is arranged in the lower portion, and vacuum suction is performed on another wall surface. A container 1 having an opening 13 communicating with the pump 12 and an opening 14O fixed to the upper surface of the container 1
A firing mechanism 2 for firing a nylon bullet having particles carrying a gene as a substance to be introduced, and a member 30 fixed to the left and right inner walls of the container 1 and having a support portion 3 at the center.
1 is arranged on the stop plate supporting device 3 having a small cross-sectional area and the supporting part 31 of the stop plate supporting device 3 to receive the nylon bullets fired from the firing mechanism 2 and to introduce the introduced substance through the hole 41. Stop plate 4 to fly to the lower sample T
And the sample T mounted in the stop plate 4 and the vacuum chamber 10.
And a filter member 5 which is disposed between and and captures dust scattered when the bullet hits the stop plate.

The container 1 is composed of a hollow rectangular parallelepiped member 10 and a front cover member 11 which is openable and closable. The container 1 is opened when a sample or nylon bullet is put in or taken out, and is otherwise airtight. It consists of a closed structure. A sample shelf 15 on which the sample T is placed is arranged at a position slightly apart from the bottom of the lower portion of the container 1. A container 16 having a trapezoidal cross section having an opening 13 formed in an approximately center of a side wall on the right side in the drawing and having an inclined surface on the front face arranged adjacently
A pipe 12P provided with a valve 12V by a suction pump 12 driven by an electric motor 12M arranged inside
The inside of the container 1 is sucked through the via to maintain the inside of the vacuum chamfer 10 in a vacuum state.

The firing mechanism 2 includes a vacuum cylinder 21 fixed to the upper surface of the container 1, a barrel 22 coaxially arranged in the vacuum cylinder 21, and an anesthesia gun coaxially arranged above the barrel 22. It is composed of a gene gun 23 composed of an equivalent special launching device. 35S / GU in the gene gun 23
Diameter 1 to 5 carrying S gene and carrying plasmid
The head is filled with micron tungsten or gold particles, and a nylon ammunition with a diameter of 5.6 mm and a gunpowder are loaded, and the nylon ammunition passes through the barrel 22 by the explosion of the gunpowder, and the upper surface of the container 1 It is configured to inject into the container 1 through the opening 14O.

The stop plate supporting device 3 is constituted by a cross-shaped member 30 having a cross-sectional area of one-tenth or less of the cross-sectional area of the vacuum chamber in which the arm is composed of four rectangular members.
Four arms 3 with flat sides on both sides at 90 degree intervals
2A are arranged radially, and are fixed to the inner wall of the container 1 at the left and right ends at a position approximately one-third above the container 1 and for fixing the stop plate 4 to the center. Support part 3
1 is arranged coaxially with the barrel 22 of the firing mechanism 2.

The stop plate 4 is the support portion 3 of the stop plate support device 3.
A metal annular plate 40 fixed to 1 is provided, and a hole 41 having a diameter of 1 mm is coaxially formed in the center.

The filter member 5 constitutes a filter device for a substance introducing device, and comprises four rectangular arms 32A and a supporting portion 31.
And a mesh member 50 composed of a mesh having a size that allows only particles carrying genes, which are introduced substances, to be arranged in four trapezoidal spaces formed between the inner wall of the container 1 and The mesh member 50 captures the dust that scatters when the bullet collides with the stop plate 4, and prevents the lower area in the vacuum chamber 10 in which the sample T is placed from being contaminated with the dust.

In the apparatus of the first embodiment having the above structure, the gene gun 23 of the firing mechanism 2 turns 35S into tungsten particles.
A nylon bullet carrying a plasmid incorporating the / GUS gene passes through the barrel 22 by the explosion of gunpowder and is fired to collide with the stop plate 4 supported by the stop plate support device 3. The stop plate 4 causes the particles carrying the gene, which is the introduced substance, to be maintained in a vacuum state in the vacuum chamber through the holes 41 and fly through the space with little resistance toward the sample T, which is a young leaf of corn, and the sample T When some of the fine particles of the carrier reach the cell and break into the cell wall along with the substance fragment to enter the cell, and stop there and release the substance fragment, the substance is introduced into the sample T.

The debris produced by the collision of the nylon bullets with the stop plate 4 and the debris produced by the explosion of the explosive fly along the strong gas flow produced by the combustion of the explosive and fly downward from above the stop plate support device 3. It runs and is captured by the mesh member 50.

The substance introduction device of the first embodiment having the above-mentioned action has an effect that the 35S / GUS gene can be introduced into the cells of the sample T in an appropriate state.

When the nylon ammunition is fired between the four arms 32A, the strong flow of gas generated by the combustion of the explosive is not blocked by the arms 32A each having a small surface area and uniformly distributed along the flat surfaces thereof. Since the gas is guided so as to flow radially downward, the flow in the vacuum chamber 10 is not biased, and there is no bias in how the introduced substance scatters, and the sample can be uniformly and uniformly reached.

Further, in the filter device for the substance introducing device of the first embodiment, the mesh member 50 causes the strong flow of the gas generated by the combustion of the explosive to get on the dust generated by the collision of the nylon bullet with the stop plate 4 or the like. The effect of trapping and reaching the sample T is obtained.

Furthermore, in the filter device for the substance introducing device of the first embodiment, since each arm 32A has a small cross-sectional area, the strong flow of gas generated by the combustion of the explosive at the time of the firing of the nylon ammunition is not obstructed. Since it is guided so as to flow radially downward between the flat surfaces on both sides of the arm 32A, the flow in the vacuum chamber 10 is uniform and uniform, and the scattering of other particles is also uniform. There is an effect that the capturing efficiency of the member 50 is high.

Further, the area surrounded by the mesh member 50 on which the sample T is placed has the effect of being able to maintain the state in which it is not contaminated by the above-mentioned dust.

(Second Embodiment) In the substance introducing apparatus of the second embodiment, the ring member 34R at the upper end is screwed to the upper wall of the container 1 having the opening 14O as shown in FIG.
The stop plate support device 34 is configured by disposing a support portion 34S for supporting the stop plate 4 at the center of a disc member 34C at the lower end suspended by four vertical members 35 fixed to 4R.

The filter device 51 includes an upper end ring 34R of a stop plate supporting device 34 and a disc member 34 above and below the mesh material.
By being locked to C, it is surrounded by a cylindrical mesh member 52, and the right half on the front side is openable and closable so as to remove the nylon bullets stopped by the stop plate 4 to form a door portion 53. In FIG. 2, the door portion 53 is in a state where the hook is removed and the door portion 53 is opened.

The device of the second embodiment having the above-mentioned structure has the same effect as that of the first embodiment, and in addition, by opening the door portion 53, the fired nylon bullet can be easily removed and the sample T There is an effect that it is possible to take a wide area to be placed, which is not polluted, and the contamination degree can be reduced.

(Third Embodiment) In the substance introducing apparatus of the third embodiment, the stop plate supporting device 36 is composed of a substantially I-shaped member as shown in FIG. One end is horizontally fixed to the inner wall of the container 1 at the left and right ends, and the support portion 37 for fixing the stop plate 4 to the center of the rectangular portion connecting the left and right ends is the barrel 22 of the firing mechanism 2. It is arranged coaxially with.

Dust cover 5C constituting the filter device
Consists of a hollow cylindrical body 55 having an inner diameter larger than the sample T and a height several times higher than the sample T at both ends, and the lower end opening 57 is arranged coaxially with the sample T on the sample shelf 15.

Mesh member 5D constituting the filter device
Is a circular member composed of a mesh of a size that captures debris and thread-like debris from the hole 41 of the stop plate 4 and allows the introduced substance to permeate therethrough, and has a circular area with a small area of about 1 mm in the center. The stopper 58 is arranged coaxially with the hole 41 of the stopper plate and placed on the upper end of the dustproof cover 5C so as to cover the opening 56.

Since the mesh member 5D is arranged on the dustproof cover 5C placed below the stop plate 4 in the apparatus of the third embodiment having the above-mentioned structure, the sample T cannot reach without passing through the mesh member 5D. Dust cover 5C and mesh member 5D
The area surrounded by has the effect that pollution is effectively prevented.

Since a soft resin nylon bullet collides with the conventional metal stop plate 4 at a high speed, the hole 41 of the stop plate 4 is formed.
Nylon bullets are scraped off at the edges of the, and thread-shaped resin pieces pop out, clogging or damaging the mesh of the mesh member, or if there is no mesh, collide with the sample cells and crush the cells. there were. In the third embodiment, since the thread-like resin fragments which may be 3 to 5 mm scattered from the hole portion 41 of the stop plate 4 can be collision-locked by the locking portion 58, the nylon bullet is The present invention has an effect of preventing dusts and thread-like debris generated upon collision with the stop plate from reaching the sample T, and preventing damage, sometimes destruction and death of cells of the sample.

Originally, the central part of the sample (immediately below the injection port)
In the cells of, the particles are likely to concentrate and cells are likely to die, and it is difficult to obtain the recombinant, but since the filamentous resin fragments have a length (may be 3 to 5 mm),
It sometimes destroyed cells other than the central part. If the sample is cells cultured in a plastic petri dish,
The thread-like resin sometimes destroyed the petri dish.

In the third embodiment, the filamentous resin collides with or comes into contact with the stop portion 58 and decelerates, so that the mesh and cells are not directly hit, and the effect of the collision is suppressed. Moreover, the particles for the locking portion 58 do not hit the central portion of the sample, but as described above, it is a portion where it is difficult to obtain a recombinant, and there is no actual damage.

(Fourth Embodiment) As shown in FIG. 4, the substance introducing apparatus of the fourth embodiment has an upper ring 3 on the upper wall 14 of the container 1.
The circular lower plate 3 is made up of four vertically arranged rod-shaped legs 39L which are screwed to the 9R and fixed to the upper ring 39R.
9P is hung coaxially, and the support portion 39S of the stop plate 4 is disposed in the center of the lower plate 39P to provide the stop plate support device 39.
Make up.

The side wall 59S and the lower wall 59B are fixed to the upper wall 14 of the container 1 and the whole of the side wall 59S and the lower wall 59B are made of the same mesh member as in the second embodiment, and a locking portion having a diameter of 1 mm is formed at the center of the lower wall 59B. The filter member 59 provided with 59K is coaxially arranged outside the stop plate supporting device 39. The front half of the filter member 59 can be opened and closed to remove nylon bullets and accumulated debris.

The substance introducing apparatus of the fourth embodiment having the above-mentioned structure has the same effects as those of the third embodiment, and the sample T
As a result, it is possible to take a large area where the slabs are not contaminated and to reduce the degree of contamination.

The embodiments described above are merely examples for the purpose of explanation, and the present invention is not limited to them. Those skilled in the art will recognize from the claims, the detailed description of the invention and the description of the drawings. Modifications and additions can be made without departing from the technical idea of the present invention.

In the above embodiments, the mesh member was described as the filter device, but the present invention is not limited to them, and a filter means having the same effect can be adopted.

In the above-mentioned embodiment, the container is a rectangular parallelepiped, but in the present invention, the flow in the container is made uniform,
From the viewpoint of eliminating the stagnation area, it is possible to employ a hollow cylindrical body or other container.

In the above-mentioned embodiments, an example in which the annular plate is used as the stop plate has been described. However, in the present invention, in order to increase the fixing area and obtain strong fixation, the outer shape such as a square or rectangle is rectangular. It is also possible to adopt the stop plate of.

As an object of the sample of the present invention, an example of a young leaf of corn was explained in the above-mentioned example, but there is no limitation in the target cell type, from a huge cell type of the cell wall to a soft type, It is applicable not only to plant cells and certain types of microorganisms, but also to microorganisms without cell walls and animal cells.

The introduced substance is, for example, 35S / GUS.
Although the example of carrying the gene on the tungsten particles loaded with the gene has been described in the examples, the introduced substance used in the present invention is not limited to this, and other genes include the dyes anthracene and sedative. Genes related to the production of useful substances such as scopolamine, dwarfing genes that control dwarfing of plants, and genes that change the morphology of the gene for luciferase, an enzyme that controls luminescence from fireflies, BT insect gene and coat of TMV -A gene that protects a plant such as a protein-encoding gene or a plasmid that incorporates these genes can be used.

In addition to tungsten particles, gold particles, ceramic particles, glass beads, latex particles and the like can be used as the particles carrying the gene.

As other introduced substances, capsules containing cytoplasm derived from other cells or physiologically active substances, particles for inspection / analysis such as magnetic fine particles and radioactive fine particles, and the like can be adopted.

[Brief description of drawings]

FIG. 1 is a partially cutaway perspective view showing a device according to a first embodiment of the present invention.

FIG. 2 is a perspective view showing a second embodiment device of the present invention.

FIG. 3 is a partially cutaway perspective view showing a device according to a third embodiment of the present invention.

FIG. 4 is a perspective view showing a device according to a fourth embodiment of the present invention.

FIG. 5 is a perspective view showing a conventional device.

[Explanation of symbols]

 1 Container 2 Firing Mechanism 3 Stop Plate Support Device 4 Stop Plate 5 Filter Member 10 Vacuum Chamber 12 Vacuum Suction Pump 13, 14 Opening 21 Vacuum Cylinder 22 Barrel 23 Gene Gun 31 Support Portion 41 Hole Portion 50 Mesh Member 58 Locking Portion

Claims (3)

[Claims] 1. A stop plate for receiving a piece of a projecting material, which is supported by a stop plate supporting device and is ejected from a firing mechanism, in the vacuum chamber of the substance introducing device, and ejects the introduced substance toward the sample. , A filter member that is placed between the sample placed in the vacuum chamber and the sample that captures the debris and debris generated when the bullet collides with the stop plate, and prevents it from reaching the sample A filter device for a substance introduction device, which is characterized by 2. The filter member according to claim 1, wherein the filter member is composed of a mesh member arranged below the stop plate and above the sample, and has a locking portion having a small area in the center thereof. A filter device for a substance-introducing device, comprising a structure in which thread-like fragments generated by collision of a bullet piece with a stop plate are collision-locked. 3. A container which forms a vacuum chamber, has an opening, and in which a sample into which a substance is introduced is placed, and a firing mechanism which ejects a piece of bullet containing the introduced substance from the opening,
It consists of a stop plate support device that has a support part that is spaced from the sample that is fixed to the container and placed in the vacuum chamber, and a member that has a hole in the center that is located in the support part of the stop plate support device. There is a stop plate that receives the projectile ejected from the firing mechanism, and on the other hand causes the introduced substance to fly toward the sample through the hole, and a filter member arranged between the stop plate support device and the sample. And a filter device configured to filter out dust and the like generated when the bullet hits the sample plate so as not to reach the sample.