TW200909579A - Low pressure accelerated gene delivery device and barrel structure thereof - Google Patents

Abstract

A barrel structure of a low pressure accelerated gene delivery device including a pressurized chamber and a spray nozzle is described. The spray nozzle is connected to the pressurized chamber, wherein the inside diameters of the spray nozzle front-terminal and the pressurized chamber are the same, and the outside diameters of the spray nozzle and the pressurized chamber are the same. The spray nozzle includes an interior contour having a converging part, a diverging part and a throat part between the converging part and the diverging part. The converging part is extending from the spray nozzle front-terminal to the throat part and the diverging part is extending from the throat part to the spray nozzle back-terminal. The connection between the converging part and the pressurized chamber is curve-shaped. The diverging part linear-divergently extends to the back-terminal of the spray nozzle. The diverging part can also parabolic-divergently extend to the back-terminal of the spray nozzle.

Description

200909579 Low-pressure airflow accelerated gene delivery device and a device with a structure that is early and easy to scale and has a transfer efficiency and its barrel tube. [Prior Art]

With the development of genetic material and the rapid development of genetics and genetics, human beings have been able to change the genetic material of cells and other organisms. With the advancement of biotechnology, 'will turn m kind _= difficult acid (DNA, deQX division. Dirty 1eic is widely used in basic research to change biological miscellaneous, has been improved by nine and Chen industry crops, such as increase Farmers; anti-candle and change the nutrients of beans, etc. In recent years, the technology of the colonization has also begun to be used

IX. INSTRUCTIONS OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to its barrel structure, and particularly to reducing gas turbulence, having a high & configuration. The soil has included genetic hiding and genetic vaccines. The successful application of the gene in the field of medical prevention and treatment will bring about major medical breakthroughs for many genetically related diseases. In recent years, the development of a gene that uses physical gene transfer technology has taken a big step forward in its application of clinicalization of gene transfer technology. In this method, a gold particle carrying a biological substance (such as DNA) is used to cause a biological substance to enter a biological cell by firing, thereby achieving the purpose of gene transfer. It has been used in many research and development fields, such as plant systems, mammalian somatic cells, gene therapy, and even the recent DNA vaccine 6 200909579 research system.

There is a new type of gene gun designed by the theory of gas dynamics, which can use a gas (such as nitrogen, helium or air) to contain a liquid solution containing biological substances (such as DNA or RNA or protein) at low pressure. Instantly accelerate to extremely high speed (greater than 2〇〇m/s), so it is not necessary to use carrying particles (such as gold, tungsten particles, etc.) to allow biological substances to pass through the surface structure or cell membrane of the organism, into the cytoplasm or nucleus. The biological cells express special foreign eggs, or gene transfer to produce new biological functions. The current application fields include animals, plants and cell lines. However, the observation of (4) the structure of the genetic test leaks has a high degree of difficulty in casting, and the phenomenon of gas turbulence is likely to occur in the storage of the gene grabbing structure, which will greatly reduce the gene transfer efficiency. SUMMARY OF THE INVENTION The purpose of the invention is to provide a low-pressure airflow acceleration device and a pipe-sucking structure which are characterized by easy praying and which have the effect of generating gas turbulence in the storage. - Construction of low-pressure airflow accelerated gene delivery | Set of barrels = structure, which includes animals and nozzles. The nozzle mouth includes the nozzle front end spot nozzle connected to the inside of the nozzle front end (4) storage =: and ===, gradually === and the pressure accumulating end is arc-shaped == 5 5200909579 The relationship between the radius G is the other The relationship between the radius of curvature Rt of the throat and the radius Ύ of the throat is < Rt < 2rr. Moreover, the gradual extension extends from the throat in a straight line to the rear end of the nozzle, wherein the diameter of the gradual extension is an increasing function, as follows:

DD^rT+(rE- rT)x/LD where x is the position of the gradual extension from the throat, Dd is the radius at the position X of the gradual extension, LD is the length of the gradual extension, and rT is the throat Radius, rE is the radius of the back end of the nozzle. In one embodiment, the throat is designed as a parallel short straight section to improve the atomization of the sample liquid. In one embodiment, the angle Θ between the fading portion of the inner contour and the center axis of the nozzle is greater than the twist and less than 15 degrees. In one embodiment, the tapered portion is a point where the straightening of the pressure accumulating chamber is linearly tapered to the position of the throat, and the diameter of the tapered portion is a decreasing function, and the linear function is: Z) c=r/-fr /- rT)x/Lc where x is the position of the tapered portion from the pressure storage tank, Dc is the radius at the position X of the tapered portion, Lc is the length of the tapered portion, rT is the radius of the throat, and η is The radius of the front end of the nozzle. The present invention further provides a barrel structure of a low pressure gas flow accelerated gene delivery device comprising a pressure storage chamber and a nozzle. The nozzle includes a front end of the nozzle and a rear end of the nozzle, and the front end of the nozzle is connected with the accumulator chamber, wherein the inner diameter of the front end of the nozzle is the same as the inner diameter of the 6 200909579 accumulator chamber, and the nozzle has an internal contour and the accumulator nozzle has an internal contour, and the inner contour includes The tapered portion, the gorge in the tapered portion and the progressive-shaped throat, wherein the tapered portion of the mouth is extended to the throat, and the progressive portion extends from the throat to the rear end of the nozzle. The relationship between the part and the accumulative end is connected by a circle, and the relationship between the half-heart of the front end of the curvature clock of the arc is 仏~. Further, the relationship between the curvature R of the Kaki and the radius π of the throat is rr < Rt <>. The drought + gradual extension is the extension of the red bell-shaped curve ship from the throat, and the exponential function of the bell curve of the gradual extension is as follows: , /, medium r = rT + b (le^) a = ~ 1 / L ^ ^lHrE-rT)/b]

b>r£~rT where x is the radius at the position of the gradual portion from the throat, and the length of the gradual extension is the radius of the apex of the nozzle, and the radius is 5 The radius of the part, the curvature calculated according to the value of the value, and the parameter of the type, β is designed according to the straight mouth of the improved mouth. In an embodiment, the μ, +, . to the throat position are connected. Lu = shrinkage is the self-accumulation chamber with a linear gradual function as ... · & her part diameter is a decreasing function, its linear

Dc=r1-(rI-rT)x/ic The position of the constricted portion, Dc is the gradual decrease, which is the self-recharged smashing end, the spurt is the second half of the throat, the radius is half, the Lc is the gradual radius The length of the part, Γτ rI is the radius of the front end of the nozzle. The invention further proposes a low-pressure airflow accelerated gene delivery device, which comprises a barrel structure, a firing device, a gas supply county, and a grabbing structure such as Weiwei. The firing device package draws and opens the device. (4) 暇 Configure the limb between the training and the cabin. The gas device is connected to the device. (4) External filament @(四)嘴 = The nozzle is firmly connected to the firing device. In one embodiment, the apparatus further includes an annular spacer disposed at the rear end of the nozzle, wherein the annular spacer has at least one gap. In an embodiment, the apparatus further includes a feed aperture located between the throat and the button. The feed hole can be delivered by biological material, metal particulate biological material or mixed with liquid and metal particles. Biological material. In one embodiment, the apparatus described above further includes a feeding device at the feed opening. ,

In one embodiment, the device control valve described above is a solenoid valve or a mechanical valve. Based on the above, the nozzle has the same outer diameter as the outer diameter of the pressure accumulator, so that the snap-off structure is easy to cast. In addition, the inner diameter of the joint between the nozzle front end and the accumulator chamber is the same as the inner diameter of the accumulator chamber, and the tapered portion and the accumulator chamber end are connected by a circular arc, thereby effectively eliminating the gas generated by entering the accumulator chamber. Spoiler phenomenon. In addition, the parallel short straight section of the throat is designed to enhance the atomization effect in addition to the sample. In order to achieve an even exit velocity, the internal profile of the progressive section is designed as a bell. 6 6200909579 On the other hand, the annular spacer is placed at the rear end of the nozzle to maintain a fixed distance between the low-pressure airflow-accelerated gene delivery device and the target to be fired, so as to reduce the shooting due to the gene gun and the intended shooting. The target object = the error caused by the distance offset, and the annular spacer has at least - a gap to prevent the strong airflow from injuring the target to be fired. The above and other objects, features and advantages of the present invention will become more apparent.

It will be understood that the preferred embodiments are described below in detail with reference to the accompanying drawings. [Draft] FIG. 1 is a schematic diagram of a low-pressure airflow accelerated gene delivery device system according to an embodiment of the present invention. Fig. 2A is a schematic view showing the structure of a barrel of the present invention. Figure 3 is a partial enlarged view of the circled portion in Figure 2A. Hereinafter, please refer to FIG. 2, FIG. 2A and FIG. 3 simultaneously, and the low-pressure gas flow accelerated gene delivery device system 1 mainly includes a gas supply reservoir 1, a barrel structure, and a firing chamber. The gas supply device 102 includes a gas η], a pressure regulator 114, and a line 116, wherein the line 116 is connected between the pressure regulator 114 and the firing device 106. The gas used is supplied by a gas cylinder 112, and the gas in 112 may be nitrogen, helium or other gases. The quality of hernia is relatively small, and it does not harm cells. It is a very ideal gas, but the price is more responsible. In some corneal plants with thicker cuticles, it is still not used. However, when using a gene gun system in some biological systems that are easier to pass, the use of nitrogen is sufficient. The gas in the cylinder 112 is first passed through a pressure regulator 114 to set the desired pressure. 5 200909579 used and sprayed "4. Qisaki 22 steel or its alloy. Pan pipe Yin Yin 1〇4, which is for example aluminum alloy, stainless easy to change the easy $. For example, disposable pipe grab, with a replacement In front of the mouth, ^4=2^ inner diameter and outer diameter, the mouth is used to store the tank 122. The front end of the nozzle is the same as the front end of the 12 servant nozzle, so it can be used for feeding and arranging. The diameter is in phase with the inner diameter of the ballast chamber 122. The inner/mouth front end coffee and the accumulator inspection 122 having the same disturbance between the flow storage chamber 122 and the nozzle have the same inner diameter, and there is no such a π angle because there is no π angle. The dead angle or the rotating body enters the accumulator chamber 122 from the rolling body supply device 102 == 2: the nozzle 124 that does not have gas turbulence when entering the front end 1243 of the nozzle has the same outer diameter as the accumulator chamber 122, so the touch is simple It is easy to pray. The material used for the nozzle 124 is a bio-methane such as a green alloy, a residual steel or an alloy thereof. The nozzle m has an inner contour, and the inner rim includes a tapered portion 126, a progressive portion 128, and a throat 130 between the tapered portion 126 and the progressive portion 128. The tapered portion 126 extends from the nozzle front end 124a to the throat portion 13b, and the progressive portion 128 extends from the throat portion 130 to the nozzle rear end 124b. In addition, the pipe grabbing structure 1〇4 may further include a barrel casing 134 which is engaged with the firing device 1〇6 by the thread on the pipe casing 134 to fix the pipe grabbing structure 104 and to make the gene gun more beautiful. The firing device 106 includes a control valve 118 and an activation device 136, wherein the control valve 118 is disposed between the activation device 136 and the accumulator compartment 122. Gas 6 6 200909579 After passing through the pressure regulator 114 and passing through the line 116, the body will first be turned on, and the device 136 controls whether or not to enter the pipetting structure (10) via the control (five) 118. The control side 118 is, for example, a mechanical valve. In one embodiment, control valve 118 is controlled via a controller (not shown). When using the low-pressure airflow accelerated gene delivery device of the present invention, the sample is first dosed from the (four) port 132, which is known, for example, by any suitable feeding device (not shown) for feeding the sample, for example Feeding is carried out using a needle, pipette or automatic delivery system. The feeding device in the structure of the present invention is not limited to a single technique or device, and any currently known structural device or method can be applied. The annular spacer 131 is sleeved at the outlet of the nozzle rear end 124b to maintain a fixed spacing between the low pressure airflow accelerated gene delivery device and the target to be fired to reduce the firing rate using the low pressure airflow accelerated gene delivery device. The error due to the offset of the distance between the low pressure airflow accelerated gene delivery device and the target to be fired. The annular spacer 131 has a gap of less than 140 ” gas flowing out of the gap 14〇 to prevent strong air current from injuring the target to be fired. In operation, the pressure regulator 114 is first set to a certain appropriate pressure and the target of the low-pressure air-accelerated gene delivery device to be fired is prepared, and a sample containing biological substances (such as RNA, DNA or protein) is prepared. The trough is placed in the feeding device. At the time of shooting, the gene smashing structure rupture 106 opens the control valve 118 via a starting device 136 to allow gas to flow into the pressure accumulating chamber 122, generating a high-speed airflow to drive the sample solution, and successfully hitting the target through the nozzle 12 200909579 124. In particular, the material m is a _ pro-riding, that is, it includes at least a tapered portion ί26, a gradual portion (3), and a continuous curve of the _ squaring 4 126 and the gradual portion 128 at the _m3q 'm Money is smooth. In addition, in order to operate the lyophilized particles (four) at a higher speed, the present invention

128 can shoot the supersonic gas, and make the solution particles more I 曰 L, so the metal can carry the particles without the need for the purpose of penetration: the throat 130 of the nozzle 124 is designed with a curved entrance, as shown in Figure 3. It allows the gas to be 'distributed at its outlet' (4), so that the solution particles will be distributed evenly. It will not collect the towel in some areas, causing cell death. Moreover, the pressure of the gas at the outlet can be close to the atmospheric pressure to avoid harming the target cell tissue. 4 inches, the control structure of the low-pressure airflow accelerated gene delivery device of the present invention has the following special structural design:

(1) Designing the throat of the tapered portion, the progressive portion and the throat 130: rT<Rt<2rT gradual portion 128: 〇<θ < 15 degree progressive portion 128 is designed to have an angular expansion The angular conical tube L^ flows through the tapered portion 126 and the throat 130 and is then ejected through the diverging portion 128. In Fig. 3, Rt represents the radius of curvature of the throat 13〇, rT is the radius of the throat 130, and one is the angle between the progressive portion 128 and the central axis (the dotted line in the figure). In order to improve the atomization effect of the sample liquid, it is necessary to add a parallel short straight pipe section in the throat 13 ,, in addition to the liquid drop for the sample, it also has the effect of improving the atomization of 13 200909579. (2) The tapered portion and the accumulator chamber are connected by a circular arc, mr =1126 and the accumulator chamber 122 are connected by a circular arc, and the relationship between the radius and the front end of the nozzle is 拉 Γ 126 and The pressure accumulation should be connected by a circular arc = so that there is no dead angle between the two or (4), thus eliminating the gas turbulence (2) the rim design of the gradual extension part #是linear expansion. ^疋, the progressive portion m extends from the throat 130 in a straight line to the spray as shown in Figure 2A), and the diameter of the progressive portion 128 is an increasing function:

DD = rT + (rE - rT) x / LD where x is the position of the progressive portion 128 from the throat 130, 〇 is the radius at the position x of the progressive portion 128, and the length of the Ld^ progressive portion 128 = rT The radius of the throat 130 is printed as the radius of the exit of the nozzle 124. In another embodiment, the inner contour of the progressive portion 28 is a bell-shaped gradation, as shown in Figure 2B. That is, the progressive portion 128 extends from the throat 13〇 in a bell-shaped curve to the rear end of the nozzle 124. The exponential function of the bell curve of the progressive portion 128 is as follows: r^=rT+b (\-e 'ax) a~ -1/Ld* In [l-(rE-rr)/b] 6 6

200909579 where x is the position of the progressive portion 128 from the throat 130, the half-pinch at the position X of the progressive portion 128, the heart being the length of the progressive portion 128, and the half pinch 130' of the portion being turned into the rear end of the nozzle 124 The radius, 6 is the parameter of the profile of the 128 part of the section 'α is the curvature calculated according to the value of 6. In the basin, the larger the value of b, the smaller the slope of the curve, and the smaller the value of the decreasing straight line ^, the larger the slope of the curve is, which is closer to the bell type. ^ In addition, the inner contour of the tapered portion is linearly tapered. In a preferred embodiment, the tapered portion 126 of Figures 2A and 2B is where the straightening of the pressure accumulating chamber 122 is linearly coupled to the position of the throat 130, and the tapered portion 126 directly invades into a decreasing function ' The linear function is: rT)x/Lc, where X is the position of the tapered portion 126 from the load bank 122, Dc is the radius at the position X of the tapered portion 126, and Lc is the length of the tapered portion 126, Γτ For the throat 130 radius, Γι is the radius of the front end of the nozzle 124. In addition, the tapered portion 126 of the nozzle 124 and the progressive portion 128 can also be designed according to the following description: The pseudo-attack flow field is isentropic flow, and the ratio of the exit area Ae to the throat area At is

Ae 1 -—>_____ At 'Me 2 γ + 1 1 + Ning Shi 2 where Me is the exit Mach number (Mach number = gas flow rate / speed of sound), if the desired Mach number and Ae are selected, the throat is available The area At and the total pressure Po of the desired nozzle inlet. 15 5200909579 Ρο = Ρ l+^Me2'Υ_ΙΪ Ρ〇= ρ|ι+υΐ1Μ. V-1 1 η 2 ω j Ld (Ld is Dump loss) s It is selected because it requires a certain speed to blow the sample solution. The Mach speed of the accumulator compartment 122 can be used to obtain the desired cylinder 112 pressure. In addition, the quality of the pressure reserve must be equal to the quality of the inflow. Jia Jia machine m, - (pAV), = (pAV)〇ut

The inlet area required to flow into the accumulator compartment 122 can be obtained from the above formula. EXPERIMENT The following is an experiment using a practical operation to investigate the position of the feed hole in the feed hole. In this experiment, the sample was 16//1 CoomaSsieBlue, using a pressure of 5 psi, and the delivery was a paper. 4A is a photographic view showing the sample being thrown from the feed hole above the throat, and FIG. 4B is a photographic view showing the sample taken from the feed hole centimeters from the throat, as shown in FIG. 4C. Photograph of Figure 4D from the sample of the input hole from the throat of 2 $8 from the throat is shown as a photo taken from the feed hole of the sample of 5.0 cm from the sample from = 16 5 200909579 4Ε A photograph of a person who is thrown from a feed hole 7.5 cm from the throat. ... Please refer to Figure 4Α, 4Ε for the soil. From the results of Figure 4Α-图4Ε, the sample of the sin near the throat is sampled, and the sample ‘ ^ is more penetrating and better. The preparation process of the sample solution, depending on the preparation details of the biological substances contained, is not limited to a single technique or a procedure, and may be any known technique or method. In general, depending on the nature of the contained organism (4), the biological substance is dissolved in a suitable solution and can be obtained by appropriate treatment. Preparation of the DNA solution will construct a complete fluorescent protein expression system DN Α Α plasm p p p p p 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌 无菌Bombardment of the device. Preparation of TE aqueous solution containing PEI PEI (polyethyleneimine, Sigma P3143, 50% (w/v) in

Water) was dissolved in sterile distilled water to make a 12.5% (w/v) aqueous solution, and then diluted to 0.00001% (0.1 ng/pl) with TE buffer to prepare a TE aqueous solution containing PEI. 17 6 6200909579 傍 DNA-ΡΕΙ complex (complex) The preparation of the complex is completed by mixing the DNA with the PEI aqueous solution at a ratio of ΡΕΙΡΝΑ=1ι^1μβ. Preparation of DNA-spermidine-CaC12 complex solution Pipette 5 ul (lug/ul) of plastid DNA (pBI121, pGR 'pCG) with 20 ul of 0.1 μM spermidine in a sonic wave to mix it evenly. Add 50 ul of 2.5MCaC12 shake mix. After standing for 5 minutes, 140 ul of 70% alcohol was added and centrifuged at 12,000 rpm for 2 seconds. After careful removal of the supernatant, 140 ul of absolute alcohol (100%) was added without disrupting the DNA pellet, and centrifugation was performed at 12,000 rpm for 2 seconds. After removing the supernatant, the DNA pellet was reconstituted with 50 ul of absolute alcohol. Preparation of EGFP protein solution The purified fluorescent protein EGFP was dissolved in sterile water and directly placed in a feeding device for bombardment of a low-pressure airflow accelerated gene delivery device. The experimental design of bombarding mouse skin with low-pressure airflow accelerated gene delivery device was selected. Five-week-old C57BL6 mice were used to remove the abdominal hair of the mouse with a razor to expose the smooth skin tissue. 'The DNA aqueous solution, qNa_pei complex aqueous solution or EGFP protein solution was placed. After entering the low-pressure airflow accelerated gene delivery device, the low-pressure airflow accelerated gene delivery device bombards the exposed skin of the mouse. The skin of each mouse's abdomen can be bombarded with four hairs, and the volume of each solution is 20 μl. The mouse was sacrificed the next day after the gene bombardment: 18 6 6200909579 to remove the skin _, remove the touch _ after treatment can be directly observed with a glory microscope. The experiment results of the low-volume airflow accelerated gene delivery device and the mouse skin were directly bombarded with DN sputum solution at a pressure of 20, 30, 40 and 50 psi, respectively, and the low-pressure airflow was accelerated with an open inner diameter ltom. The bombardment of the gene delivery device, each hair, 2 〇 μιη, contains 3jIgDNA, and under the fluorescent microscope, the fluorescent performance of the skin of the abdomen bombarded by the old milk is observed. Fig. 5-8_5D shows the results of a 40-fold magnification of a fluorescent microscope, which are sequentially applied at 2, 3, 40 and 50 psi. From the experimental results, it can be seen that the skin tissue bombarded at 3 psi and 40 psi has the best green fluorescence performance. The PEI-DNA complex aqueous solution was bombarded with a helium gas at a pressure of 30 psi, and the bombardment tube with an opening diameter of 1 〇mm was used to bombard the low-flow air-accelerated gene delivery device, and each solution was a 2 μμ1 PEI-DNA complex. The aqueous solution contained (iv) and observed the fluorescent appearance of the mouse's bombarded abdomen skin under a fluorescent microscope. As shown in Figure 6, when PEI is added to the solution, the DNA dose can be reduced to

At the time of Wg, the fluorescent expression is equivalent to that of the aqueous DNA solution containing 3 DNA per hair. Bombardment with EGFP protein 19 200909579 The low-pressure gas-accelerated gene delivery device shoots with a helium gas psi pressure of '5//g EGFP protein per hair. The fluorescence of the abdominal skin of the mouse was observed under a fluorescent microscope. As shown in Fig. 7A to Fig. 7C, the abdominal skin of the mouse was observed by a fluorescence microscope 40 times magnification, and it was observed that the EGFP protein was successfully and uniformly injected into the epidermal layer. Petal Onf Oncidium Ramsey with a low-seat airflow-accelerated gene delivery device, with a helium gas pressure of 50 psi, with a barrel with an opening diameter of 4 5 mm for bombardment of a low-pressure airflow-accelerated gene delivery device. The solution was a 1 μμ1 DNA-Spermidine-CaC12 complex aqueous solution containing 1 μ§ DNA, the muzzle was 3 cm away from the target, and GUS tissue staining was performed on the next day. 'GUS was observed in the dark room for 48 hours. The coloring situation. It can be seen from the foregoing experimental results that the low-pressure airflow accelerating gene delivery device used in the present invention does not cause serious damage to the target cell tissue. Moreover, the low-pressure airflow accelerated gene delivery device of the present invention can directly transfer genetic DNA or other biological substances into the interior of the cell, thereby avoiding the consumption of barriers hindered by many tissues or cells, and successfully achieving gene transfer or biological substances. The purpose of delivery. In summary, the present invention has at least the following advantages: 1. In the low-pressure airflow accelerated gene delivery device proposed by the present invention, the nozzle of the pipe-collecting structure has the same outer diameter as that of the pressure storage tank, making the barrel structure easy. Binding. 20 6 200909579 2. The connecting device of the low-pressure airflow accelerated gene delivery device proposed by the present invention has the same inner diameter as the connection portion of the front section of the accumulator compartment, and the tapered portion and the pressure accumulating compartment end are connected by a circular arc, thereby eliminating When the gas enters the accumulator compartment, the flow is disturbed. 3. The inner contour of the gradual portion of the pipe grabbing structure of the present invention is a linear expansion or a bell-shaped expansion, and such a design can increase the acceleration of the emitted gas and provide a high uniformity of the emitted gas.

Ο 4. The low-pressure airflow accelerated gene delivery device proposed by the present invention has an annular spacer, which can keep a fixed interval between the low-pressure airflow accelerated gene delivery device and the target to be shot, so as to reduce the shooting error. In addition, since the annular door spacer of the low-pressure airflow-accelerated gene delivery device proposed by the present invention has at least one notch, it is possible to prevent the strong airflow from injuring the object to be fired. Although the present invention has been disclosed in the above preferred embodiments, it is not intended to limit the present invention, and it is to be understood that those skilled in the art can make some modifications and refinements without departing from the spirit of the invention. The scope of the invention = the scope of protection is subject to the definition of the scope of the patent application. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a low-pressure airflow acceleration delivery device according to an embodiment of the present invention. FIG. 2A and FIG. 2B are diagrams showing a pipe grabbing structure according to an embodiment of the present invention. As shown in Fig. 3, a partial enlarged view of the circled portion in Fig. 2A is shown. Figure 4B shows a photograph of a sample taken from a feed hole above the throat. Figure 21 6200909579. Figure 4B is a photograph of a sample taken from a feed hole that is 12 centimeters from the throat. Figure 4C is a photograph of a sample taken from a feed hole 2 · 5 cm from the throat. Figure 4D is a photograph of a sample taken from a feed port that is 5. centimeters from the throat. Figure 4E is a photograph showing a sample taken from a feed hole 7 cm from the throat. Fig. 5 is a magnification of 4 times in a fluorescent microscope, and the observation result of bombarding the abdominal skin of the mouse with a helium gas of 20 psi is a magnification of 40 times under a fluorescent microscope, and the observation result of the abdominal skin of the mouse is 30 psi. 5C is a 40-fold magnification under a fluorescent microscope, and the observation results obtained by hitting the abdominal skin of a mouse with 40 psi of helium. Figure 5D is a magnification of 40 times under a glory microscope, and the observation result of bombarding the abdominal skin of a mouse with 50 psi of helium is shown in Fig. 6 40 times magnification under a fluorescent microscope, the results of bombardment of the abdominal skin of mice with PEI-DNA complex water>glutle solution '30 psi of helium gas. Figure 7 A is not painted with EGFP protein crit mouse skin tissue, in fluorescence The results (non-delivery area) were observed under a microscope (magnification 40 times). Fig. 7B is a view showing that the mouse skin tissue was bombarded with EGFP protein, and the result (delivery area i) was observed under a fluorescence microscope (magnification 40 times). Figure 7C is a view showing the results of bombardment of mouse skin tissue with EGFP protein, 22 6 6200909579 under a fluorescent microscope (magnification 40 times) (the delivery area is not in the form of alcohol (no DNA), 5 psi) Observations on the staining of G. sinensis by the GUS tissue after the sputum of the scorpion (4) control) ° Figure 8B shows the observation of the GUS tissue staining after bombarding the petals of Oncidium with pBI121 solution and 5 psi of helium. result. Figure 8C is a graph showing the results of staining GUS tissue with a pRG solution and 5 psi of helium bombardment of the petals of Oncidium. In Fig. 8D, the observation results obtained by staining GUS tissue with the pCG solution and the gas of 5 psi are used. [Description of main component symbols] 100: Low-pressure airflow accelerated gene delivery device 102: Gas supply device 104: Pipe grabbing structure 106: Fire-fighting device 106 112: Cylinder 114: Pressure regulator 116: Pipe H8: Control valve 122: Ballast 124 = nozzle 124a: nozzle front end 124b: nozzle back end 200909579 126: taper 128: taper 130: throat 132: feed hole 134: pipe grab housing 136: starting device

Ae: cross-sectional area of the end of the progressive portion 128

At : throat 130 cross-sectional area

Rt: radius of curvature of the throat 130 rT: radius of the throat 130 Θ : angle between the progressive portion 128 and the central axis

L 24

Claims (1)

6 200909579 X. Patent application scope: 1. A barrel structure of a low-pressure airflow accelerated gene delivery device, comprising: a pressure storage tank; and a nozzle comprising a nozzle front end and a nozzle rear end, the nozzle front end An accumulator chamber connection, wherein an inner diameter of the front end of the nozzle is the same as an inner diameter of the accumulator chamber, the nozzle having an inner contour, wherein the inner contour comprises: a tapered portion; a gradual portion; and a throat portion Located between the tapered portion and the gradual portion, wherein the tapered portion extends from the front end of the nozzle to the throat, the gradual portion extending from the throat to the rear end of the nozzle, wherein the tapered portion The pressure storage tank end is connected by a circular arc, and the relationship between the radius of curvature of the circular arc and the radius r/ of the front end of the nozzle is 2r / 5 between the radius of curvature Rt of the throat and the radius of the throat The relationship is < Rt < 2rr, the gradual extension extends from the throat in a straight line to the rear end of the nozzle, and the diameter of the gradual extension is an increasing function, as follows: DD = rT + (rE - rT)x/LD where x is the gradation from the throat The position of the portion, Dd is the radius at the position X of the gradually increasing portion, LD is the length of the progressive portion, rT is the radius of the throat, and rE is the radius of the rear end of the nozzle. 25 5 200909579 2. The pipetting structure of the low-pressure airflow accelerated gene delivery device according to claim 1, wherein the throat is designed as a parallel short straight pipe section to improve the atomization effect of the sample liquid. ^ 3. The pipetting structure of the low-pressure airflow accelerated gene delivery device according to claim 1, wherein the angle between the gradual portion and the central axis of the nozzle is greater than the twist and less than 15 degree. 4. The pipe grabbing structure of the low-pressure airflow-accelerated gene delivery device according to the invention of claim 2, wherein the taper is tapered from the accumulator chamber to the throat, the taper portion The diameter is a decreasing function, and its linear function is: Dc=n-(ri- rT)x/Lc where X is the position of the taper from the accumulator, and Dc is the taper position. The radius at X, Lc is the length of the tapered portion, and the radius 'rl' of the throat is the radius of the front end of the nozzle. 5) a pipetting structure of a low-pressure airflow accelerated gene delivery device, comprising: - a pressure storage tank; and a mountain nozzle comprising a nozzle front end and a nozzle rear end, the nozzle being connected to the pressure storage tank, wherein The inner end of the nozzle front end is the same as the inner pressure of the pressure accumulating remainder, and the outer diameter of the nozzle is the same as the outer diameter of the pressure accumulating chamber, the nozzle has an inner contour, wherein the inner contour comprises: a tapered portion; a gradual portion; and 26 5 200909579 - a throat, recorded __ riding a tapered portion extending from the front end of the nozzle to the throat portion, wherein the extension to the rear end of the nozzle, & the progressive portion from the throat In the extension, the tapered portion and the pressure accumulating chamber end have a radius of curvature Ri of the arc and the front of the nozzle. The arc is connected, the relationship between the circle H and the thousand is Rl. The radius of curvature of the throat is the relationship between the radius of the throat and the throat is rr < Rt < 2rT, 礼1々 The throat is at the back end of a bell, and the exponential function of the bell curve of the gradual extension is extended to the jet r=rT+b (l-e'ax) a= -1/Ld* In [\~{ rE~rT)/^ j· Material ^ X f From the presumption, the ship's inspection 7 is the radius of the gradual position of the gradual extension, and the heart is the length of the gradual extension. The radius of the part, q is the radius of the back end of the nozzle, 6 is the parameter defining the shape of the gradual part, and α is the curvature calculated according to the ό value. 6. The pipetting structure of the low-pressure airflow accelerated gene delivery device according to claim 5, wherein the throat is designed as a parallel short straight pipe section to improve the atomization effect of the sample liquid. 7. The pipe grabbing structure of the low-pressure airflow-accelerated gene delivery device according to claim 5, wherein the tapered portion is a line that is tapered from the end of the accumulator chamber to the position of the throat, The diameter of the tapered portion is a decreasing function, and its linear function is: 6 200909579 rT)x/ic The taper is the position of the tapered portion from the /I storage 'ballast, DC is the fade & The radius at which U is the length of the tapered portion, f is the radius of the throat, and ri is the radius of the front end of the nozzle. T~Λ 8.: a low-pressure airflow accelerated gene delivery device, comprising: a f-superstructure, as described in claim 1 or 5 of the patent application; a transmitter device comprising: a control valve and a And the control is arranged in the New Zealand rogue recital; the first gas supply device is connected to the firing device; and a pipe grab casing is used for fixing the mouth to be turned over to the forwarding device. 4 mouth 'to make the actual delivery K application? The low-pressure gas flow-accelerating gene described in the eighth aspect of the patent range includes: - an annular spacer is placed at the rear end of the crucible, and /, the middle spacer has at least one notch. Ο The low-pressure airflow accelerating base sheet described in item 8 of the patent application scope includes a feed hole, which is disposed at the throat of the nozzle and the gradual biological B. The feed hole can be delivered. Liquid biological material, metal particles 'raw material or biological material mixed with liquid and metal particles. • The low-pressure air-accelerated base-receiving unit, as described in claim 8 of the patent application, further includes a feeding device installed at the inlet. 2. The low-pressure airflow acceleration type retransfer device as described in claim 8 wherein the control valve is a solenoid valve or a mechanical valve. 28