JPH0631496U - Stop plate for substance introduction device and substance introduction device - Google Patents

Abstract

(57) [Abstract] [Purpose] To prevent bullets from bouncing and rolling due to their elasticity when they collide with a stop plate, to position bullets and improve operability. [Structure] A circular step portion 42 coaxially formed with a hole portion 41 formed in the center of a metal annular plate 40 and a vertical portion 42V.
Stop plate 4 composed of and. Top part 1 of vacuum chamber 10
4 is provided with an opening 14O, a sample is placed in the lower part thereof, and an opening 13 is formed on the other wall surface to communicate with the vacuum suction pump 12, and a substance fixed to the upper surface of the container 1 and introduced through the opening 14O. A firing mechanism 2 for firing a nylon bullet having particles carrying genes and a member 30 fixed to the left and right inner walls of the container 1 and having a support portion 31 in the center and a stop plate supporting device having a small cross-sectional area Three
And a filter member 5 arranged between the stop plate 4 and the sample T, and the stop plate 4 arranged on the support portion 31 of the stop plate support device 3.

Description

[Detailed description of the device]

[0001]

[Industrial applications]

 According to the present invention, a bullet is collided with a stop plate supported by a stop plate supporting device in a vacuum chamber to introduce genes and other introduced substances carried by the bullet into living cells or tissues and other samples. The present invention relates to a stop plate for a substance introduction device and a substance introduction device for gripping a piece of bullet that has collided with the stop plate on a stepped portion.

[0002]

[Prior art]

 As shown in FIG. 8, a vacuum chamber C formed in a housing means H is divided into upper and lower chambers UC and DC by a horizontal portion M as shown in FIG. However, the upper and lower chambers UC and DC constitute independent chambers that are independently vacuum-controlled by conduits UL and DL having cutoff valves UV and DV that communicate with the respective surge tanks S. A collision receiving mechanism R, which has a hole in the center of the horizontal portion M and has a flat surface and is composed of a disk member having a flat surface, is arranged. The bullets that have collided with the disc member R and are covered with the substance to be released into the microbial sample from the central opening O are sent into the biological material BS that is the sample, and the multiple bullets have the above-mentioned central opening. There is one that seals O and seals the upper and lower chambers UC and DC.

[0003]

[Problems to be solved by the device]

 Since the above-mentioned conventional stop plate for substance introduction device is composed of a disk member having a flat surface, when a multi-shot piece body collides, it is rebounded by the elasticity of the multi-shot piece body and rolls on the flat surface, so the position is fixed. However, there was a problem that the operability was poor.

In the above-mentioned conventional substance introduction device, since 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, the vacuum control of the upper and lower vacuum chambers UD and DC is performed respectively. Since it is necessary to independently operate the shutoff valves UV, DV and the conduits UL, DL of the above, there is a problem that the operation of the vacuum control is difficult and the structure is complicated.

Therefore, the inventors of the present invention focused on the technical idea of having a function of locking the colliding piece with a stop plate that collides and stops the piece in the vacuum chamber, and also continued research and development. As a result, the bullet collided with the stop plate by forming a stepped portion composed of a circular flat surface that is thinner than the other portions coaxially with the hole for passing the introduced substance formed in the center. At this time, the projectile is locked by the stepped part, and its elasticity prevents it from bouncing back and rolling on a flat surface, so that the position of the projectile is fixed and the operability is improved. The present invention has been achieved that achieves the purpose of facilitating the construction and simplifying the structure.

[0006]

[Means for Solving the Problems]

 A stop plate for a substance introducing device according to the present invention (the first invention according to claim 1) is mounted in a vacuum chamber of the substance introducing device, a firing mechanism for firing a piece of a piece containing an introduced substance, and a vacuum chamber. It is a stop plate that is placed between the sample and the sample that is held by the stop plate support device and that receives the projectiles with the introduced substance that have been launched from the firing mechanism and flies the introduced substance toward the sample. It is configured such that a step portion formed of a circular flat surface that is thinner than the other portions is formed coaxially with the formed hole for passing the introduced substance.

The stopper plate for a substance introducing device according to the present invention (the second invention according to claim 2) is the stopper plate according to the first invention, wherein the diameter of the portion near the circular flat surface of the step is the step stop plate. The structure is formed to have a diameter larger than that of the portion close to the upper surface of the.

The substance introducing apparatus of the present invention (the third invention according to claim 3) forms a vacuum chamber, has an opening, and a container for placing a sample into which the substance is introduced, and an introducing substance from the opening. A firing mechanism for firing the provided bullet pieces, a stop plate supporting device in which a supporting part is arranged at an interval with respect to a sample fixed in a container and placed in a vacuum chamber, and a supporting part of the stopping plate supporting device. It is a member that has a hole formed in the center where it is placed and a step formed coaxially with the hole that is composed of a circular flat surface that is thinner than other parts. On the other hand, a stop plate that receives the other substance and causes the introduced substance to fly toward the sample through the hole is formed.

[0009]

[Action]

 The stopper plate for a substance introducing device according to the first aspect of the present invention having the above-described structure is configured such that when a bullet piece containing the introduced substance emitted from the launch mechanism of the substance introducing device collides with the stopper plate, the bullet piece is engaged by the step portion. It stops and prevents it from bouncing due to the elasticity of the bullet and rolling on a flat surface.

In addition to the function of the first invention, the stopper plate for a substance introducing device of the second invention having the above-mentioned structure has a diameter of a portion near the circular flat surface of the step portion on the upper surface of the stop plate of the step portion. Since it is formed larger than the diameter of the close part, the step of locking the bullet piece effectively acts to grasp and lock the bullet piece, and the elasticity of the bullet piece causes it to bounce off and roll on a flat surface. It is one layer of effective prevention.

In the substance introducing device of the third invention having the above-mentioned structure, the projecting mechanism ejects the bullet piece provided with the introducing substance, and the bullet piece is placed in the vacuum chamber by the supporting portion of the stop plate supporting device. When it hits, the projectile is locked by the stepped part to prevent it from bouncing due to the elasticity of the projectile and rolling on a flat surface, and at the same time, particles carrying the substance fly toward the sample through the hole. Then, the substance is introduced into the sample.

[0012]

[Effect of device]

 In the stopper plate for the substance introducing device of the first invention that has the above-mentioned action, since the bullet piece is locked by the step portion of the stopper plate, the position of the bullet piece on the stop plate is determined and the ejected bullet piece is removed. This has the effect of making the operation easier.

In addition to the effect of the first aspect of the invention, the stop plate for the substance introducing device of the second aspect of the present invention, which has the above-described function, effectively holds the bullet pieces by the stepped portions having different diameters. The position of the bullet on the stop plate is more effectively determined, and the operation of removing the fired bullet becomes easier.

In the substance introducing device of the third invention having the above-mentioned action, when the bullet piece collides with the stop plate, the bullet piece is locked by the step portion, so that the position of the bullet piece on the stop plate is determined and the bullet piece is fired. In addition to the effect of facilitating the operation of removing the fragments, the particle carrying the substance is made to fly toward the sample through the hole to introduce the substance into the sample and to Since there is only one vacuum chamber, the operation of vacuum control of the vacuum chamber is simple, and the structure is simple.

The substance introduction device of the third invention having the above-mentioned action achieves the effect of the first invention, and at the same time, arrives at debris and thread-like fragments passing through the hole of the stop plate and other processes. Since the dust is captured and blocked by the filter member of the filter device, it is possible to introduce only the necessary substance into the sample.

[0016]

【Example】

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

First Embodiment As shown in FIGS. 1 to 3, the first embodiment relates to a stop plate 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. The following is simply referred to as a substance introduction device.

As an example, the substance introduction device of the first embodiment allows a sample T of a young leaf slice of maize, which is the second leaf after one week of germination, to carry a plasmid incorporating the 35S / GUS substance on tungsten particles. It is a thing to shoot with a thing and introduce.

The stop plate 4 for the substance introducing device of the first embodiment is arranged on the support portion 31 of the stop plate supporting device 3 fixed in the container 1 described later and fixed to the support portion 31 of the stop plate supporting device 3. As shown in FIGS. 1 and 2, it is composed of an annular plate 40 made of metal and having an outer diameter of 25 mm and a thickness of 7 mm. A hole 41 having a diameter of 1 mm is coaxially formed in the center of the annular plate 40, and a circular step portion 42 having a diameter of 8 mm and a depth of 1 mm is formed coaxially with the hole 41. The ejected nylon bullet is received by the circular flat surface 42B, locked by the vertical portion 42V, and particles carrying the introduced substance are made to fly to the lower sample T through the hole 41 formed at the center. The hole portion 41 is configured such that the diameter of the lower portion is gradually expanded so that the particles carrying the introduced substance fly to the entire sample T.

As shown in FIGS. 1 to 3, the substance introducing apparatus of the first embodiment forms a vacuum chamber 10, forms an opening 14O in the upper surface portion 14, arranges a sample in the lower portion, and A container 1 having a wall formed with an opening 13 communicating with a vacuum suction pump 12, and a firing mechanism 2 for firing a nylon bullet having a particle carrying a gene as an introduced substance from an opening 14O fixed to the upper surface of the container 1. A stop plate supporting device 3 fixed to the left and right inner walls of the container 1 and having a supporting part 31 in the center and a small cross-sectional area; and a supporting part 31 of the stopping plate supporting device 3. The stop plate 4 and the filter member 5 that is disposed between the stop plate 4 and the sample T placed in the vacuum chamber 10 and captures the dust scattered when the projectile collides with the stop plate 4. Become.

The container 1 is composed of a hollow rectangular parallelepiped member 10, and the front part thereof is composed of a lid member 11 which can be opened and closed. The container 1 is opened when a sample or a nylon bullet is put in or taken out, and is otherwise opened. It consists of a structure that closes tightly. 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 valve 12V is operated by a suction pump 12 driven by an electric motor 12M arranged in a container 16 having a trapezoidal cross section having an inclined surface on the front face and an opening 13 formed in the approximate center of the right side wall in the figure. The inside of the container 1 is sucked through the provided pipe 12P to maintain the inside of the vacuum chamber 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 inside the vacuum cylinder 21, and an anesthesia gun coaxially arranged above the barrel 22. The material gun 23 is composed of an equivalent special launching device. The gun is filled with 35S / GUS material in the material gun 23, and the head is filled with tungsten or gold particles having a diameter of 1 to 5 micron and carrying a plasmid. It is configured such that a nylon bullet is loaded, passes through the barrel 22 by the explosion of gunpowder, and is injected into the container 1 through the opening 14O on the upper surface of the container 1.

The stop plate supporting device 3 is constituted by a cross-shaped member 30 having a cross-sectional area which is one-tenth or less of the cross-sectional area of the vacuum chamber in which the arm is composed of four rectangular members, and is arranged at intervals of 90 degrees. It has four arms 32A radially formed on both sides, and is fixed horizontally to the inner wall of the container 1 at left and right ends at a position approximately one-third above the container 1. Further, a support portion 31 for fixing the stop plate 4 is disposed in the center coaxially with the barrel 22 of the firing mechanism 2.

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

In the device of the first embodiment having the above-mentioned structure, the nylon gun carrying the plasmid in which the 35S / GUS substance is incorporated in the tungsten particles is passed through the barrel 22 by the explosion of the gunpowder by the substance gun 23 of the firing mechanism 2. It is fired and collided with the stop plate 4 supported by the stop plate support device 3.

The stop plate 4 collides with the flat surface 42B of the circular step portion 42 to lock the side wall of the laterally expanded nylon bullet with the vertical portion 42V, and to make a hole for the particle carrying the gene as the substance to be introduced. A space with a low resistance maintained in a vacuum chamber in the vacuum chamber is made to fly toward the sample T, which is a young leaf of corn, through the portion 41, and reaches the sample T. 35S 2 / GUS, which is the substance to be introduced, is introduced into the sample T by breaking the cell wall and invading into the cell, stopping there and releasing the substance fragment.

The dust generated by the collision of the nylon bullet with the stop plate 4 and the dust generated by the explosion of the explosive fly on the strong flow of the gas generated by the combustion of the explosive and fly downward from above the stop plate supporting device 3. It runs and is captured by the mesh member 50.

The stopper plate for the substance introducing device of the first embodiment having the above-mentioned action hits the flat surface 42B of the circular step portion 42 and laterally spreads the nylon bullets by the vertical portion 42V to lock and bounce. Since it prevents backing and rolling and stabilizes the position, it has the effect of good operability.

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

When a nylon bullet is fired between the four arms 32A, since each arm 32A has a small surface area, it does not interfere with the strong flow of gas generated by the combustion of explosives, and along the flat surfaces thereof. Since the particles are guided so as to flow radially downward uniformly, there is no bias in the flow in the vacuum chamber 10 and there is no bias in the scattering of the particles carrying the substance, and it is possible to reach the sample T uniformly and uniformly. Produce an effect.

In addition, 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, and the scrap generated by the collision of the nylon bullet with the stop plate 4 or the like. Of the mesh member 50 to prevent the mesh member 50 from reaching the sample T, and since the scattering of dust and the like is also uniform, the efficiency of capturing the mesh member 50 is high.

Further, the area in which the sample T surrounded by the mesh member 50 is placed has an effect that it is possible to maintain a state in which the sample T is not contaminated.

(Second Embodiment) The stop plate 4 of the second embodiment is an embodiment of the second invention, and has substantially the same configuration as that of the first embodiment as shown in FIGS. The point is that the vertical inner wall of the circular step portion 42 has a large inner diameter in the concave portion 42D close to the flat bottom surface 42B of the circular step portion 42, and the shoulder portion 42S projects inward and is formed in the small inner diameter. It is that you are.

In the stop plate 4 of the second embodiment having the above structure, the nylon bullets fired from the firing mechanism collide with the flat bottom surface 42 B of the circular step portion 42, and the side wall of the nylon bullet drops the circular step portion 42. The small-diameter shoulder portion 42S protruding when coming into contact with the inner wall directly bites into the side wall of the nylon bullet, and the lower portion of the nylon bullet enters the recess 42D, so that the nylon bullet is reliably locked.

(Third Embodiment) As shown in FIG. 6, the substance introducing apparatus of the third embodiment includes four upper rings 39R screwed to the upper wall 14 of the container 1 and fixed to the upper ring 39. The circular lower plate 39P is coaxially suspended by the rod-shaped leg portions 39L arranged vertically, and the support portion 39S of the stop plate 4 is arranged at the center of the lower plate 39P to form the stop plate support device 39.

As shown in FIG. 7, the stop plate 4 supported by the support portion 39S of the stop plate support device 39 has a hole portion 41 having a diameter of 1 mm at the center of the metal annular plate 40 as in the above-described embodiment. Is formed coaxially, and a circular step portion 42 having a diameter of 8 mm and a depth of 1 mm is formed coaxially with the hole portion 41. The inner side wall of the circular step portion 42 is formed by a tapered surface 42T, and is formed so that the wedge-shaped projection 42P and the gap 42K are formed such that the inner diameter increases as it approaches the lower flat bottom surface 42B.

A filter member 59 fixed to the upper wall 14 of the container 1 and having the side wall 59S and the lower wall 59B wholly formed of a mesh member and having a locking portion 59K with a diameter of 1 mm arranged at the center of the lower wall 59B. Are arranged coaxially outside the stop plate support device 39. The filter member 59 has a front half that can be opened and closed so that nylon bullets and accumulated debris can be removed.

In the substance introducing apparatus of the third embodiment having the above-mentioned structure, the nylon bullets fired from the firing mechanism collide with the flat bottom surface 42 B of the circular step portion 42, and the side wall of the nylon bullet has the circular step portion 42. When the inner wall is in contact with the inner wall, the inner wall is composed of the taper surface 42T and the wedge-shaped protrusion 42P, so that the wedge action causes the nylon bullet to enter and wedge into the wedge-shaped gap 42K, and the wedge-shaped protrusion 42P. By digging into the side wall of the nylon bullet, the nylon bullet can be securely locked.

Further, in the apparatus of the third embodiment, since the side wall 59S and the lower wall 59B are entirely composed of the mesh member so as to surround the stop plate 4, the sample T cannot be reached without the mesh member. In addition to the function and effect, the area outside the mesh members 59S and 59B is effectively prevented from being contaminated.

In the third embodiment, the thread-like resin shards 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 59K. With this, it is possible to prevent the scraps and thread-like debris generated when the nylon bullets collide with the stop plate from reaching the sample T, and prevent the cells of the sample T from being damaged, sometimes destroyed, and killed. Has the effect.

Furthermore, the third embodiment has an effect that the uncontaminated area where the sample T is placed can be widened and the contamination degree can be reduced.

The embodiments described above are merely examples for the purpose of explanation, and the present invention is not limited to them, and it will be understood by those skilled in the art from the claims of utility model, the detailed description of the invention and the description of the drawings. Modifications and additions can be made as long as they do not violate the technical idea of the structure of the present invention that can be recognized by.

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

In the above-mentioned embodiments, the mesh member has been described as the filter device, but the present invention is not limited thereto, and a filter means having the same effect can be adopted.

In the above-mentioned embodiment, the container is a rectangular parallelepiped, but as the present invention, a hollow cylindrical body or other container is adopted from the viewpoint of making the flow in the container uniform and eliminating the stagnation area. Is possible.

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

Further, the example in which the substance to be introduced and the particles are also carried on the tungsten particles in which the 35 / GUS substance is incorporated has been described in the example, but the present invention is not limited to this, and other than that, gene Examples include genes related to the production of useful substances such as the pigment anthracene and the analgesic scopolamine, dwarfing genes that control plant dwarfing, and genes that encode proteins of the luciferase enzyme that controls firefly-derived luminescence. Genes that protect plants and plasmids that incorporate these genes can be used.As particles that carry genes, in addition to Tungsten and gold particles, ceramic particles, glass beads, latex particles, etc. can be used. is there.

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 radiation fine particles, and the like can be adopted.

[Brief description of drawings]

FIG. 1 is a perspective view showing a stop plate according to a first embodiment of the present invention.

FIG. 2 is a vertical sectional view showing a stop plate of the first embodiment.

FIG. 3 is a partially cutaway perspective view showing the substance introducing apparatus of the first embodiment.

FIG. 4 is a perspective view showing a stop plate according to a second embodiment of the present invention.

FIG. 5 is a vertical sectional view showing a stop plate according to a second embodiment.

FIG. 6 is a perspective view showing a substance introducing device according to a second embodiment.

FIG. 7 is a vertical sectional view showing a stop plate according to a third embodiment of the present invention.

FIG. 8 is a perspective view showing a conventional device.

[Explanation of symbols]

 1 Container 2 Firing Mechanism 3 Stop Plate Supporting Device 4 Stop Plate 5 Filter Member 10 Vacuum Chamber 12 Vacuum Suction Pump 13, 14 Opening 21 Vacuum Cylinder 22 Barrel 23 Material Gun 31 Support 40 Ring Member 41 Hole 42 Circular Step 42B Circular Flat surface 42D Recessed portion 42K Gap 42P Projection 42S Shoulder portion 42T Tapered surface 42V Vertical portion 50 Mesh member 59K Locking portion

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Ichiro Takakura Inventor 1-2-1 Yokou, Kawasaki-ku, Kawasaki-shi, Kanagawa Nihon Zeon Co., Ltd. Research and Development Center (72) Tatsuo Numasuzawa Yokou, Kawasaki-ku, Kawasaki-shi, Kanagawa 2-1 Nichihon Zeon Co., Ltd. Research and Development Center

Claims (3)

[Scope of utility model registration request] 1. A vacuum plate of the substance introduction device, wherein a stop plate supporting device is arranged between a firing mechanism that fires a piece of bullet containing the introduction substance and a sample placed in the vacuum chamber. It is a stop plate that receives the projectiles with the introduced substance that is fired from the device and flies the introduced substance toward the sample.The stop plate is coaxial with the hole formed in the center for passing the introduced substance. A stopper plate for a substance introducing device, characterized in that a step portion constituted by a circular flat surface that is thinner than the portion is formed. 2. The substance introducing device according to claim 1, wherein a diameter of a portion of the step near the circular flat surface is larger than a diameter of a portion of the step near the top surface of the stop plate. Stop plate. 3. A container which forms a vacuum chamber and has an opening, on which a sample for introducing a substance is placed, a firing mechanism for ejecting a piece of bullet containing the introduction substance through the opening, and a vacuum chamber fixed to the container. A stop plate supporting device in which a supporting part is arranged at a distance to the sample placed inside, and a hole is formed in the center of the supporting part of the stopping plate supporting device and is coaxial with the hole part. It is a member that has a stepped portion composed of a circular flat surface that is thinner than other parts.It receives a piece of bullet fired from the firing mechanism, and on the other hand, the introduced substance flies toward the sample through the hole. A substance introduction device comprising: a stop plate for running.