CN1284383A - Ejecting pneumatic gene accelerating microejection method and gene gun - Google Patents
Ejecting pneumatic gene accelerating microejection method and gene gun Download PDFInfo
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- CN1284383A CN1284383A CN00122801A CN00122801A CN1284383A CN 1284383 A CN1284383 A CN 1284383A CN 00122801 A CN00122801 A CN 00122801A CN 00122801 A CN00122801 A CN 00122801A CN 1284383 A CN1284383 A CN 1284383A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
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Abstract
The present invention belongs to the field of biological technology. High-pressure air is first accelerated to form supersonic airflow, which is accelerated for the second time for ever high speed, and between the first and the second acceleration, one negative pressure port is formed to led micro pellet in for acceleration together with the airflow so as to reach required speed to shoot target cell. The method of the present invention can accelerate micro pellet to speed high enough to shoot different target cell, and the preparation of gene micro pellet is simple without need of specific equipment.
Description
The invention belongs to biological technical field, particularly be used for gene vaccine (claiming dna vaccination again) is imported human or animal's the epidermis or the special equipment of histiocytic method and this method at body.
Gene vaccine is a vaccine of new generation, and origin comes from antigen encoding gene of pathogen and forms as the plasmid DNA of genophore.This section antigen encoding gene can be controlled synthetic antigen albumen in active somatic cell, thereby cause immunoreation, be on molecular level, virus to be suppressed, make it inactivation, improve body immunity, be that with the difference of common vaccine the antigen protein of gene vaccine is to produce in the body of immune object.Gene vaccine has been opened the gate that leads to new immunization route.How with the gene vaccine transfered cell, and obtaining effective expression, is one of key technology of performance gene vaccine effect.Traditional method is directly the saline solution of naked DNA to be injected in the animal body with syringe, cause damage by muscle or skin histology to the injection site,, injecting anesthetic medicine freezing as the part or snake venom make tissue be in reproduced state, just can further improve the conversion ratio of DNA.This method consumption is big, and transfection efficiency is low, also has misery, thereby suddenly waits to improve.U.S. BIO-RAD company has several years ago released a kind of hand-held particle gun, its structure and operation principle as shown in Figure 1, the helium in the high pressure helium source of the gas 1 is contained in the plastic tubulature 4 that shoots in the dish 3 by shooting dish seat 2, entering.Little bullet 5 of wrapping up in gene is adsorbed on the inwall of short tube 4.After gases at high pressure enter short tube 4, little bullet 5 is blown away, enter accelerated passage 6.After little bullet 5 quickens, remove the gunnery target cell through tapering trumpet-like passage 8 again in accelerated passage 6.Regulating part 9 both can be located, and can adjust shooting distance again; O type circle 7 seals, and prevents gas leakage.The advantage of this structure is that little bullet places in the passage of gases at high pressure, is easy to quicken and disperse.The shortcoming that exists is that preparation is complicated, needs special equipment earlier little bullet to be adsorbed on the tubule inwall, is cut into full-length again.Simultaneously, because the turbulent flow wall attachment effect of tube wall, often little bullet can not be blown away entirely, and is residual in addition.
The objective of the invention is to overcome the shortcoming and defect of above-mentioned technology, little bullet method of the pneumatic accelerated gene of a kind of novel injection type and particle gun thereof are provided.This method not only can accelerate to sufficiently high speed with little bullet, satisfies the requirement of the different target cells of shooting, and the little bullet preparation of gene is simple, need not special equipment.
The present invention proposes the little bullet method of the pneumatic accelerated gene of a kind of injection type, it is characterized in that: may further comprise the steps:
(1) gases at high pressure is carried out quickening the first time earlier, make it to become supersonic airstream;
(2) again this supersonic airstream is carried out the second time and quicken, make it further to improve speed;
(3) between quickening for the first time and quickening for the second time, radially forming a negative pressure mouth, so that little bullet is introduced from the negative pressure mouth;
(4) little bullet is introduced the back from the negative pressure mouth and is quickened with the air-flow that quickens for the second time, removes the gunnery target cell after making it to reach required speed.
The present invention proposes a kind of pneumatic particle gun of injection type that adopts above-mentioned accelerated method, comprise high-pressure air source, gas flow controller, accelerator, little bullet, little missile-borne valut and location and range adjustment unit, it is characterized in that, the injection accelerator that said accelerator is made up of the two-stage valve that fuses, the inlet of close this secondary valve has a radial negative pressure mouth; Said little bullet is arranged on this negative pressure mouth place, introduces in the said secondary valve by negative pressure.
Said one-level valve inlet can be cylindrical hole, and inlet diameter can be 3～8mm, and outlet can be circular cone, and plating degree can be 1: 10～and 1: 50, the minimum-value aperture can be 2～6mm, and transition between the two can be the accelerated passage of Wei Duoxinsi base curves formation; Going into, exporting of this secondary valve all can be taper hole; This negative pressure hole diameter can be 1～3mm.
Can be the negative pressure channel of taper seat between said one-level valve and the secondary valve, the gap can be 0.1～0.3mm.
Said little bullet can be adsorbed on the fine-structure mesh of the little missile-borne sheet of sieve shape, and said slide glass can be placed on discoid little missile-borne valut.
Said target position localizer can be the small circle ring of being convenient to the supersonic airstream diffusion of side symmetry perforate.
Said gas flow controller can comprise the pressure adjusting pressuring valve of selecting high-pressure air source output as required, the electromagnetic valve of control gases at high pressure through and off and the control circuit of control electromagnetic valve make-and-break time.
But said time controller can be made up of start button, time selector switch, intervalometer and electrical switch.
The said slide library of the present invention once can be placed multi-disc, can select arbitrarily according to design, the slide glass that is loaded with the little bullet of required emission be positioned at exactly wait to penetrate on the position (over against and be close to the negative pressure suction inlet).
The pneumatic little bullet accelerated method of this injection type provided by the invention and be exclusively used in the particle gun of this method, it can be done in gymnastics, and the golden microgranule of wrapping up in gene is quickened, and makes it to obtain enough kinetic energy, and the gunnery target cell is with the exogenous gene transfered cell.The present invention can adjust the size of air pressure and Firing Velocity and the shooting time that duration of ventilation is controlled little bullet by the air-flow control unit, satisfy the requirement of different target cells, only need little bullet is adsorbed on the sieve shape slide glass, need not special equipment, simple to operate, have flexible, easy and the efficient advantages of higher.
Brief Description Of Drawings:
Fig. 1 is existing pneumatic particle gun structural principle sketch map.
Fig. 2 is a kind of embodiment population structure sketch map of the present invention.
Fig. 3 is the pneumatic little bullet acceleration principle sketch map of injection type of the present invention.
Fig. 4 is the structural representation of the little missile-borne valut of present embodiment.
Fig. 5 is the little missile-borne chip architecture of a present embodiment sketch map.
Fig. 6 is a present embodiment electromagnetic valve make-and-break time control circuit schematic diagram.
Embodiment to the pneumatic particle gun of injection type that adopts the pneumatic little bullet accelerated method of injection type of the present invention is illustrated below in conjunction with accompanying drawing.
Present embodiment is made up of high-pressure air source, air-flow control unit, injection accelerator, little bullet, little missile-borne valut and location and range adjustment unit, as shown in Figure 2.Wherein, high-pressure air source 10 is the high-pressure helium gas cylinders on the market, and the air-flow control unit is by pressure regulator valve 11, and electromagnetic valve 14 and time controller 25 are formed, and the injection accelerator is by one-level valve 15, O type circle 17, and secondary valve 18, negative pressure valve 19 and little missile-borne sheet 21 are formed; Little missile-borne valut is formed (concrete structure as shown in Figure 4) by slide library disc seat 16 and slide library disk 22 etc., and location and range adjustment unit are made up of range adjustment part 23 and localizer 24.The annexation of each parts is: high-pressure air source 10 links to each other with pressure regulator valve 11, pressure regulator valve 11 joins with electromagnetic valve 14 by high-pressure air pipe 12 and pipe joint 13, electromagnetic valve 14 links to each other with time controller 25 and controlled by it, one-level valve 15 directly and electromagnetic valve 14 join, the secondary valve links to each other with the one-level valve, range adjustment part 23 links to each other with secondary valve 18, localizer 24 links to each other with range adjustment part 23, little missile-borne valut links to each other with one-level valve 15, and being loaded with needs little missile-borne sheet 21 of the little bullet 20 of emission to be positioned on the position of being close to negative pressure valve 19.
The implementation method of present embodiment is: little bullet 20 of wrapping up in gene that will prepare is adsorbed on little missile-borne sheet 21, is placed in the slide library disk 22, be positioned at negative pressure valve 19 directly over and be close to.With target position localizer 24 the embodiments of the invention particle gun is positioned at and shoots on the target position, regulate pressure regulator valve 11, the output gases at high pressure are transferred on the needed pressure, time controller 25 was chosen on the needed time, press the start button (see figure 6), electromagnetic valve 14 is opened, and gases at high pressure enter one-level valve 15.Gases at high pressure are accelerated into supersonic airstream by behind the one-level valve 15, enter secondary valve 18, produce negative pressure at negative pressure valve 19.Be adsorbed on little bullet 20 on little missile-borne sheet 21 under the effect of negative pressure, break away from slide glass 20, introduce from negative pressure valve 19, by the negative pressure channel between one-level valve 15 and the secondary valve 18, enter the accelerated passage of secondary valve 18, be accelerated with supersonic airstream, remove the gunnery target cell after reaching the speed of predetermined ideal.Regulate pressure regulator valve 11, can select the gases at high pressure of different pressures, make little bullet 20 obtain different speed, satisfy the shooting requirement of different target cells.Select the different make-and-break time of electromagnetic valve 14 by time controller 25, can improve fire effect.Little missile-borne valut once can be placed 6-12 slide glass, rotates slide glass disk 22, can select needed slide glass arbitrarily, and it is positioned at above the negative pressure valve 19.Shooting is once rotated one time disk 22, changes a kind of slide glass, can continuous fire.Regulate localizer 24, can change the shooting distance of little bullet 20, adjustable range can be 10-30mm, obtains different fire effects.
The present embodiment each several part is formed and function is described in detail as follows respectively:
The injection accelerator of present embodiment as shown in Figure 3, it is by one-level valve 15, O type circle 17, secondary valve 18 and negative pressure valve 19 are formed.Secondary valve 18 links to each other with one-level valve 15, and O type circle 17 seals between the two-stage valve.During work, the little missile-borne sheet that adsorbs little bullet 20 is attached on the negative pressure valve 19.Its operation principle is, the inlet of one-level valve 15 is a circular hole, diameter can be 3-8mm, after gases at high pressure enter one-level valve 15, the accelerated passage part after-acceleration that is shaped as the Wei Duoxinsi base curves by wherein becomes supersonic airstream, can be 1 through tapering: 10-1: 50 and the lowest calibre taper hole that can be 2-6mm enter secondary valve 18.After entering secondary valve 18, negative pressure interruption-forming negative pressure at inlet, negative pressure mouth diameter can be 1-3mm, little missile-borne sheet 21 is pasting negative pressure valve 19, the little bullet 20 that is adsorbed on the slide glass 21 is inhaled under the effect of negative pressure, after the gap can be the negative pressure channel of 0.1-0.3mm between one-level valve and the secondary valve, the accelerated passage that enters secondary valve 18 quickened.
Little missile-borne valut of present embodiment as shown in Figure 4, it is made up of little missile-borne valut pedestal 16, spring 26, steel ball 27, minor axis 28, nut 29, pad 30, disk 22, screw 31, pad 32, making leaf spring 33 and little missile-borne sheet 21.Slide library pedestal 16 is installed on the one-level valve 15, and spring 26 is installed in the aperture of slide glass pedestal 16, and steel ball 27 is placed on the spring 26.Minor axis 28 is installed on the slide library pedestal 16, and disk 22 is installed on the minor axis 28, and pad 30 is installed on the disk 22 by minor axis 28, and nut 29 is installed on the minor axis 28, is used to regulate the end play of disk 22, guarantees to rotate flexible, steady.Little missile-borne sheet 21 is placed in the stepped hole of disk 22, and a plurality of stepped holes when design (specifically by fixed) are arranged on the disk 22, once can place a plurality of little missile-borne sheets.Pad 32 and making leaf spring 33 are installed on the disk 22 by screw 31, and making leaf spring 33 is pushed down little missile-borne sheet 21, makes it motionless.Making leaf spring 33 can rotate, and is convenient to the loading and unloading of little missile-borne sheet 21, adjusts the thickness of pad 32, can change the pressure of stage clip 33.A plurality of fossettes (corresponding with the little missile-borne sheet number that can place) are arranged on the stepped hole aperture face of disk 22, and each recess is all corresponding with a stepped hole, i.e. the position of a corresponding little missile-borne sheet.When disk 22 is installed, steel ball 27 is just in time embedded in the fossette, adjust spring 26, disk 22 is rotated flexibly, guarantee that again steel ball 27 can play the role of positioning.Rotary disk 22 as required, and armed little missile-borne sheet 21 is positioned on the launching workplace.
The structure of little missile-borne sheet 21 of present embodiment as shown in Figure 5, it is made up of big circular slice 34, fine-structure mesh 35 and sequin 36, all has circular hole, fine-structure mesh 35 to be placed between big circular slice 34 and the sequin 36 in the middle of two kinds of disks, the three is a stainless steel material entirely, is welded into one.
Present embodiment air-flow time controller realizes that with hardware as shown in Figure 6, but it is made up of start button, time selector switch, intervalometer, electrical switch.The selection of time switch links to each other with intervalometer, and intervalometer was set on the needed time; Start button links to each other with intervalometer, starts intervalometer work; Intervalometer links to each other with electrical switch, the work of control electrical switch.During work, by the selected required time of selection of time switch, press start button, a signal is exported in intervalometer work, opens electrical switch, connects the power supply of electromagnetic valve 14, and electromagnetic valve 14 is opened gases at high pressure and flow through, and enters one-level valve 15.When timing arrives, intervalometer sends a signal again, closes electrical switch, cuts off the power supply of electromagnetic valve 15, closed electromagnetic valve, and high pressure draught cuts off.
Claims (8)
1, the little bullet method of the pneumatic accelerated gene of a kind of injection type is characterized in that: may further comprise the steps:
(1) gases at high pressure is carried out quickening the first time earlier, make it to become supersonic airstream;
(2) again this supersonic airstream is carried out the second time and quicken, make it further to improve speed;
(3) between quickening for the first time and quickening for the second time, radially forming a negative pressure mouth, so that little bullet is introduced from the negative pressure mouth;
(4) little bullet is introduced the back from the negative pressure mouth and is quickened with the air-flow that quickens for the second time, removes the gunnery target cell after making it to reach required speed.
2, a kind of employing is according to the pneumatic particle gun of the injection type of the described accelerated method of claim 1, comprise high-pressure air source, gas flow controller, accelerator, little bullet and location and range adjustment unit, it is characterized in that, the injection accelerator that said accelerator is made up of the two-stage valve that fuses, the inlet of close this secondary valve has a radial negative pressure mouth; Said little bullet is arranged on this negative pressure mouth place, introduces in the said secondary valve by negative pressure.
3, according to the pneumatic particle gun of the described injection type of claim 2, it is characterized in that, said one-level valve inlet is cylindrical hole, inlet diameter is 3～8mm, outlet is circular cone, the plating degree is 1: 10～1: 50, and the minimum-value aperture is 2～6mm, the accelerated passage that transition between the two constitutes for the Wei Duoxinsi base curves; Going into, exporting of this secondary valve all is taper hole; This negative pressure hole diameter is 1～3mm.
4, according to the pneumatic particle gun of the described injection type of claim 2, it is characterized in that, is the negative pressure channel of taper seat between said one-level valve and the secondary valve, and the gap is 0.1～0.3mm.
According to the pneumatic particle gun of the described injection type of claim 2, it is characterized in that 5, said little bullet is adsorbed on the fine-structure mesh of the little missile-borne sheet of sieve shape, said slide glass is placed on discoid little missile-borne valut.
According to the pneumatic particle gun of the described injection type of claim 2, it is characterized in that 6, said target position localizer is the small circle ring of being convenient to the supersonic airstream diffusion of side symmetry perforate.
7, according to the pneumatic particle gun of the described injection type of claim 2, it is characterized in that said gas flow controller comprises the pressure adjusting pressuring valve of selecting high-pressure air source output as required, the electromagnetic valve of control gases at high pressure through and off and the control circuit of control electromagnetic valve make-and-break time.
According to the pneumatic particle gun of the described injection type of claim 7, it is characterized in that 8, said time controller is made up of start button, selection of time switch, intervalometer and electrical switch.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001228013A CN1180847C (en) | 2000-08-25 | 2000-08-25 | Ejecting pneumatic gene accelerating microejection method and gene gun |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001228013A CN1180847C (en) | 2000-08-25 | 2000-08-25 | Ejecting pneumatic gene accelerating microejection method and gene gun |
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| Publication Number | Publication Date |
|---|---|
| CN1284383A true CN1284383A (en) | 2001-02-21 |
| CN1180847C CN1180847C (en) | 2004-12-22 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB001228013A Expired - Fee Related CN1180847C (en) | 2000-08-25 | 2000-08-25 | Ejecting pneumatic gene accelerating microejection method and gene gun |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7547292B2 (en) | 2001-01-11 | 2009-06-16 | Powderject Research Limited | Needleless syringe |
| CN105936876A (en) * | 2016-07-08 | 2016-09-14 | 魔水科技（北京）有限公司 | Water mist carrier gene gun |
| CN113091532A (en) * | 2021-03-11 | 2021-07-09 | 哈尔滨工程大学 | Compound driven second grade projectile body accelerating device and experiment platform |
-
2000
- 2000-08-25 CN CNB001228013A patent/CN1180847C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7547292B2 (en) | 2001-01-11 | 2009-06-16 | Powderject Research Limited | Needleless syringe |
| USRE43824E1 (en) | 2001-01-11 | 2012-11-20 | Powder Pharmaceuticals Inc. | Needleless syringe |
| CN105936876A (en) * | 2016-07-08 | 2016-09-14 | 魔水科技（北京）有限公司 | Water mist carrier gene gun |
| CN105936876B (en) * | 2016-07-08 | 2018-08-17 | 魔水科技（北京）有限公司 | Water mist vector gene rifle |
| CN113091532A (en) * | 2021-03-11 | 2021-07-09 | 哈尔滨工程大学 | Compound driven second grade projectile body accelerating device and experiment platform |
| CN113091532B (en) * | 2021-03-11 | 2021-09-21 | 哈尔滨工程大学 | Compound driven second grade projectile body accelerating device and experiment platform |
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| Publication number | Publication date |
|---|---|
| CN1180847C (en) | 2004-12-22 |
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