TW200909579A - Low pressure accelerated gene delivery device and barrel structure thereof - Google Patents
Low pressure accelerated gene delivery device and barrel structure thereof Download PDFInfo
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200909579 種低壓氣流加速式基因投遞裝置及 是·有關於一種構造間早而容易鱗造且 因轉殖效能的裴置及其槍管結 【先前技術】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]
隨著基因物質的發 掌握運用基因遺傳技^見與遺傳學之快速發展,人類已能 至於-個生物個體鮮=射卜來基因去改變—細胞、乃 他種生物的基因物質^表現。隨著生物科技的進步’將 _轉m種_=難酸(DNA,deQX师。臟1eic 廣泛地運用在基礎研藉以改變生物雜狀,已被 =九與辰業作物的改良上,例如增加農 几;、抗烛性和改變豆營養成分等等。近年來, ,殖的技術也開始被利用二^二疾二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
九、發明說明: 【發明所屬之技術領域】 本發明是有關於 其槍管結構,且特別 可降低氣體擾流,具有高& 構。 土 ’已括基因躲與基因疫苗等。而基因猶於醫藥預防 ,治療領域的成功應用,將為許多與遺傳相關之疾病,帶 來醫療上的重大突破。 近年來,新開發出使用物理性的基因轉殖技術之基因 搶’其已使基因轉殖技術臨床化之應用,向前邁進一大步。 該方法是利用攜帶著生物性物質(如·· DNA)的金粒,以 射擊的方式而使生物性物質進入生物細胞,達到基因轉殖 的目的。目前已應用於許多研究開發的領域,如植物系統、 哺乳動物的體細胞、基因治療,乃至於最近的DNA疫苗 6 200909579 研究系統。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.
目前有一種利用氣體動力學之理論所設計的新型基因 槍,可在低壓下,以氣體(如氮氣、氦氣或空氣等)將含 有生物性物質(如DNA或RNA或蛋白質等)的液體溶液 瞬間加速至極高速度(大於2〇〇 m/s),故無須使用攜帶微 粒(如金、鎢微粒子等)即可使生物性物質穿過生物體的 表層結構或細胞膜,進入細胞質或細胞核中,而使生物細 胞表現特殊外來蛋自質,或基因轉殖而產生新的生物功 能,目前應用領域包含動物、植物及細胞株。 然而’目摘使㈣基因檢的結構漏造成鑄造困難 性高’而对基因搶之搶管結構的蓄壓射容易產生氣體 擾流現象,會大幅降低基因轉殖效能。 【發明内容】 美ΐ發明的目的是提供一種低壓氣流加速式 裝置及其搶管結構,其具有容易禱造的特性,且 效嶋在蓄中產生氣體擾流現象。 -構種低壓氣流加速式基因投遞|置的槍管 =構,其包括畜以及喷嘴。嘴嘴包括噴嘴前端斑喷嘴 接’其中喷嘴前端的内㈣蓄 =:與===,漸=== 與蓄壓搶端以圓弧相銜== 5 5200909579 端的半徑G之間的關係為另外,喉部的曲率半徑 Rt與喉部之半徑Ύ之間的關係為< Rt < 2rr。而且,漸 張部為自喉部處以直線擴張延伸至喷嘴後端,其中漸張部 直徑為漸增函數,如下: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 其中x為自喉部算起的漸張部的位置,Dd為漸張部位 置X處的半徑,LD為漸張部的長度,rT為喉部的半徑,rE 為喷嘴後端的半徑。 在一實施例中,上述之喉部為平行短直管段設計,以 改善樣品液體之霧化效果。 在一實施例中,上述之内部輪廓中漸張部與噴嘴之中 心轴之夾角Θ大於〇度且小於15度。 在一實施例中,上述之漸縮部為自蓄壓艙以直線漸縮 至喉部位置相銜接之處,漸縮部直徑為漸減函數,其線性 函數為: Z)c=r/-fr/- rT)x/Lc 其中x為自蓄壓艙算起之漸縮部的位置,Dc為漸縮部 位置X處的半徑,Lc為漸縮部的長度,rT為喉部半徑,η 為喷嘴前端的半徑。 本發明另提供一種低壓氣流加速式基因投遞裝置的槍 管結構,其包括蓄壓艙以及喷嘴。喷嘴包括噴嘴前端與喷 嘴後端,喷嘴前端與蓄壓艙連接,其中喷嘴前端的内徑與 6 200909579 蓄壓艙的内徑相同,且喷嘴的特與蓄壓艙 噴嘴具有内部輪廓,内部輪廓包括漸縮部、峽 於漸縮部與漸張狀_喉部,其中漸縮部嘴= 伸至喉部,漸張部自喉部延伸至噴嘴後端。] 部與蓄驗端以圓_銜接,且圓弧之曲率钟 前端的半心之間的關係為仏~。另外,崎的曲= 徑Rt與喉部之半徑π之間的關係為rr<Rt<>。旱+ 漸張部為自喉部紅鐘型曲線舰延伸 r而^ 漸張部的鐘型曲線之指數函數如下: 、 /、中 r=rT+b (l-e^) a=~1/L^^lHrE-rT)/b]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 其中x為自喉部算起之漸張部的位置 置X處的半徑,忍為漸張部的長 為漸張雜 為噴嘴後端的半徑,5為定羞々為喉部的半徑、 據办值所計算的曲率、科型的參數,β則是依 改善之嘴部為平値直管段設計’以 在一實施例中,μ、+、. 至喉部位置相銜接之卢=縮部為自蓄壓艙以直線漸縮 函數為·· &她部直徑為漸減函數,其線性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 縮部的位置,Dc為漸縮 其為自蓄麼搶端算起之漸 200909579 為喉部半;, 部位置χ處的半徑,Lc為漸縮部的長度,Γτ rI為喷嘴前端的半徑。 本發明另提出-種低壓氣流加速式基因投遞裝置,盆 包括槍管結構、擊發裝置、氣體供縣置以及搶 ^ 搶管結構如総魏。擊發裝置包抽制似及啟ς裝 置’其中㈣暇配置肢練置與艙之間。氣體^ 應裝置與科裝置連接。㈣外狀絲@㈣嘴 = 喷嘴得以穩固地與擊發裝置銜接。 +在一實施例中,上述之裝置更包括環狀間隔物,套置 於噴嘴後端處,其中環狀間隔物具有至少一缺口。 在-實施例中,上述之裝置更包括進料孔,進料孔位 於喉部與紐部之間’進料孔可投遞有賴生物性材料、 金屬微粒生物性材料或是混有液體與金屬微粒的生物性材 料。 在一實施例中,上述之裝置更包括進料裝置, 進料口處。 、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. ,
在一實施例中,上述之裝置控制閥為電磁閥或機械閥。 基於上述,噴嘴具有與蓄壓艙外徑相同的外徑,因此 搶官結構容易進行鑄造。此外,喷嘴前端與蓄壓艙連接處 之内徑與蓄壓艙的内徑相同,且漸縮部與蓄壓艙端以圓弧 相銜接,因此能有效地消除氣體進入蓄壓艙所產生的擾流 現象。另外,喉部的平行短直管段設計,除用於放置樣品外, 更可加強霧化效果。而為使出口速度達到均勻,漸張部内部輪 廓得設計為鐘型。 6 6200909579 另一方面,環狀間隔物套置於噴嘴後端,可使得低壓 氣流加速式基因投遞裝置與欲射擊的目標物之間保持固^ 間距,以降低進行射擊時由於基因槍與欲射擊的目標物= 間的距離偏移所產生的誤差,且環狀間隔物具有至少—缺 口,以防止強力氣流對欲射擊的目標物產生傷害。 、 為讓本發明之上述和其他目的、特徵和優點能更明顯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.
易懂,下文特舉較佳實施例,並配合所附圖式,作詳細說 明如下。 、D 【實施方式】 圖1所繪示為本發明一實施例之低壓氣流加速式基因 投遞裝置系統的示意圖。圖2A所繪示為本發明—實施例 之槍管結構的示意圖。圖3所緣示為圖2A中圈示處的局 部放大圖。以下,請同時參照圖i、圖2A及圖3,低壓氣 流加速式基因投遞裝置系統1〇〇主要包括氣體供庫 1〇2、槍管結構刚及擊發襄置廳。 氣體供應裝置102包括氣報η]、壓力調整器114及 管路116,其中管路116連接於壓力調整器114與擊發裝 置106之間。所使用之氣體是由氣瓶112所供應的, 112内的氣體可為氮氣、氦氣或其它氣體。氦氣的質量較 小,較不傷細胞,是相當理想的氣體,但是價格較責;在 某些角質層較厚的動植物,仍是非使用氦氣不可。不過, 當使用基因槍系統在一些較容易穿過的生物系統,利用氮 氣就足夠了。氣瓶112的氣體先經過一個壓力調整器114, 以設定所需壓力。 5 200909579 所使用及喷”4。崎ί22 鋼或其合金。搶管尹槿1〇4 、,其例如是鋁合金、不銹 換容易的優$。例如是可拋棄式搶管,具有替 括嘴嘴前^4=2^内徑與外徑,嗔嘴以包 蓄麼艙122。喷嘴前端„的12仆’噴嘴前端以a連結於 同,因此可以哺广^·辨* &、内徑與畜壓艙122的内徑相 流現象。流蓄髮舱122與喷嘴間所產生的擾 相同的内/ 嘴前端咖與蓄壓驗122具有 116具有相同的内徑,因 1出π 角存在,ϋ此M h 1者連處不會有死角或轉 田體自軋體供應裝置102進入蓄壓艙122 ==2:進入嗔嘴前端1243時便不會有氣體擾流的 噴嘴124具有與蓄壓艙122相同的外徑,因此 觸簡單而容易進行禱造。喷嘴124所使用之材 科為生物性姆金屬,例如綠合金、残鋼或其合金等。 Ο 喷嘴m具有内部輪廓,内部輪扉包括漸縮部126、 漸張部128及位於漸縮部126與漸張部128之間的喉部 130。漸縮部126自噴嘴前端124a延伸至喉部13〇,漸張 部128自喉部130延伸至喷嘴後端124b。 此外,搶管結構1〇4更可包括一槍管外殼134,藉由 搶管外殼134上的螺紋與擊發裝置1〇6銜接,以固定搶管 結構104,且可使得基因槍更加美觀。 擊發裝置106包括控制閥118及啟動裝置136,其中 控制閥118是配置於啟動裝置136與蓄壓艙122之間。氣 6 6 200909579 體在經過壓力調整器114的並經過管路116之後’會先由 啟,裝置136控制是否經由控制㈤118進入搶管結構⑽ 的蓄壓搶122巾。而此控侧118例如是f賴或機械閥。 在一實施例中,控制閥118是經由一控制器(未繪示)所控 制。 卫 當於使用本發明之低壓氣流加速式基因投遞裝置時, 會先將樣本從㈣口 132投人,其例如是已知任何適 用的進料裝置(未繪示)將樣品進行送料,例如是利用針 頭、移液管(pipette)或是自動投遞系統進行送料。本發明基 因知的結構中之進料裝置不限於單一技術或装置,可以應 用任何目前已知之結構裝置或方法。 環狀間隔物131是套置於喷嘴後端124b出口處,可使 得低壓氣流加速式基因投遞裝置與欲射擊的目標物之間保 持固定間距,以降低利用低壓氣流加速式基因投遞裝置進 行射擊時由於低壓氣流加速式基因投遞裝置與欲射擊的目 標物之間的距離偏移所產生的誤差。環狀間隔物131具有 ,少一缺口 140 ’氣體可從缺口 14〇處流出,以防止強力 氣流對欲射擊的目標物產生傷害。 操作時,先將壓力調整器114設定到某—合適的壓 力’並將低壓氣流加速式基因投遞裝置欲射擊的目標物準 備好,把含有生物性物質(如RNA、DNA或蛋白質)之 樣本’谷液放入進料裝置内。射擊時,基因搶結構上之擊發 裂置106經由一啟動裝置136開啟控制閥118,使氣體流 入該蓄壓艙122内,產生高速氣流帶動樣本溶液,經喷嘴 12 200909579 124順利擊中目標物。 特別是,料m乃料為_親騎 即其至少包括漸縮部ί26、漸張部⑶以及位於縮^ _m3q ’m之目_漸縮 4 126與漸張部128之_成連續曲線,以錢體 暢。另外,為使溶賴粒㈣更高速運行,本發明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可射射氣體達到超音速,而使溶液微粒更 I曰L,故無須金屬攜帶微粒,即可達到貫穿射入之目 的:噴嘴124的喉部130設計有曲線入口,如圖3所示, 可讓氣體在其出口處’分布較㈣,進而使溶液微粒會分 佈車乂均勻’不會集巾在某些區域,造成細胞死亡的現象。 而且’可使氣體在出口處之壓力,接近―大氣壓,而避免 傷害目標細胞組織。 4寸別是,本發明之低壓氣流加速式基因投遞裝置的搶 管結構具有下列特殊結構設計: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:
(一)有關漸縮部、漸張部及喉部的設計 喉部 130 : rT<Rt<2rT 漸張部128 : 〇<θ < 15度 漸張部128的設計為一具有㊀角度擴張角的錐形管 L^氣流通過漸縮部126及喉部130後,隨即通過漸張 部128射出。圖3中Rt代表喉部13〇的曲率半徑,rT則為 喉部130的半徑,而㊀則是漸張部128與中心軸(圖中虛 線)的夾角。為改善樣品液體之霧化效果,得在喉部13〇 增設一平行短直管段,除可供樣品液體懸滴,亦兼具改善 13 200909579 霧化之效。 (二)漸縮部與蓄壓艙以圓弧相銜接 mr =1126與蓄壓艙122之間是以圓弧相銜接,且此 與噴嘴前端的半徑~之間的關係為拉 二Γ 126與蓄壓請之間以圓弧相銜接 =使免兩者之間沒有死角或㈣,因而消除氣體擾流現 ( (二)漸張部的輪靡設計 ^例中’漸張部128之内部輪#是直線擴張。 ^疋,漸張部m為自喉部130處以直線擴張延伸至喷 如圖2A所示),且漸張部128的直徑為漸增函數:(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 其中x為自喉部130算起的漸張部128的位置,〇 為漸張部128位置x處的半徑,Ld^漸張部128的長度= rT為喉部130的半徑,印為喷嘴124出口的半徑。 在另一實施例中,漸張部丨28之内部輪廓是鐘型漸 張,如圖2B所示。也就是說,漸張部128為自喉部13〇 處以一鐘型曲線擴張延伸至喷嘴124後端,漸張部128的 鐘型曲線之指數函數如下: r^=rT+b (\-e'ax) a~ -1/Ld* In [l-(rE-rr)/b] 6 6DD = 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 其中x為自喉部130算起之漸張部128的位置, 漸張部128位置X處的半捏,心為漸張部128的長度 ,部的半捏130 ’化為喷嘴124後端的半徑,6為 張部128外型的參數’ α則是依據6值所計算的曲率。盆 中,b的數值越大,曲線斜率·越小,減近直線^ 的數值越小,曲線斜率變化越大’較接近鐘型。 ^另外,有關漸縮部的内部輪廓是直線漸縮。在一較佳 實施例中,圖2A與圖2B中的漸縮部126為自蓄壓艙122 直線漸縮至喉部130位置相銜接之處,漸縮部126直 侵為一漸減函數’其線性函數為: rT)x/Lc 、其中X為自蓄壓艙122算起之漸縮部126的位置,Dc 為漸縮部126位置X處的半徑,Lc為漸縮部126的長度, Γτ為喉部130半徑,Γι為噴嘴124前端的半徑。 另外,喷嘴124的漸縮部126與漸張部128的還可以 依據以下說明來設計: 假攻流場為等熵流動(isentropic flow),出口面積Ae 與喉部面積 At比為·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 +宁施2 其中Me為出口馬赫數(馬赫數=氣體流速/音速),若 選定所需要的馬赫數及Ae,則可獲得喉部面積At以及所 需噴嘴入口之全壓Po。 15 5200909579 Ρο = Ρ l+^Me2'Υ_ΙΪ Ρ〇 = ρ|ι+υΐ1Μ. V-1 1 η 2 ω j Ld (Ld 為 Dump loss) s因需一定的速度使其能吹送樣本溶液,故選定進入蓄 壓艙122之馬赫速(Min) ’則可獲得所需氣瓶112壓力。 此外’為供蓄壓搶122内維持定壓】 出之質量需等於流入之質量。 贾賈机m, - (pAV), = (pAV)〇utAe 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
由上式即可獲得流入蓄壓艙122所需之入口面積。 實驗 以下利用實際操作的實驗,對進料孔的位置盘投遞咬 果進行研究。在此實驗中,投入的樣本為16//1的 CoomaSsieBlue,使用的壓力為5〇psi,投遞物為渡紙。 圖4A所繪示為將樣本從喉部上方的進料孔投入的照 片圖,圖4B所繪示為從將樣本從距喉部12公分的進料孔 投入的照片圖,圖4C所繪示為從將樣本從距喉部2 $公八 的進料孔投入的照片圖’圖4D所緣示為從將樣本從= 16 5 200909579 將樣 邰5.0公分的進料孔投入的照片圖,圖4Ε 本從距喉部7.5公分的進料孔投人的照片圖。 … 土請參照圖4Α,4Ε,由圖4Α-圖4Ε的結果 罪近喉部的稿孔投人樣本,樣本 ‘ ^越 且穿透性也較佳。 又禾孕乂為岣勻而 樣本溶液製備 樣本溶液之製備程序,視所含生物性物質之不 有不同之製備細節,故不限於單—技術或程序,可以 ^何目前已知之技藝或方法。—般而言,視所含生物㈣ 質之特性岐’在;={;添加金屬攜帶錄情況下,將生物性 物質溶於適當溶液中,並經適當處理即可獲得。 製備DNA溶液 將構築完整之螢光蛋白表現系統的DNΑ質體(plasmid) pEGFP-N2 >谷於無菌之TE buffer中,即可直接置於進料裝 置中’用於低壓氣流加速式基因投遞裝置之轟擊。 製備含PEI之TE水溶液 將 PEI(polyethyleneimine,Sigma P3143,50%(w/v) inThe 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)溶於無菌蒸餾水中使成為12.5%(w/v)水溶液,再以 TE buffer 稀釋為 0.00001%(0.1ng/pl),即製得含 PEI 之 TE 水溶液。 17 6 6200909579 製傍 DNA-ΡΕΙ 複合物(complex) 以ΡΕΙΡΝΑ=1ι^1μβ的比例將DNA與PEI水溶液混 合均勻即完成複合物之製備。 製備DNA-spermidine-CaC12複合物溶液 吸取 5ul (lug/ul)質體 DNA (pBI121、pGR ' pCG)與 20ul的0·1Μ spermidine於超音波震盪讓其混合均勻。再 加入50ul的2.5MCaC12震盪混合。靜置5分鐘後,再 加入140ul的70%酒精,以12000rpm轉速離心2秒。 小心去除上清液後,以不破壞DNA pellet加入140ul絕對 酒精(100%),12000rpm轉速離心2秒。除去上清液後, 以50ul絕對酒精回溶DNA pellet。 製備 EGFP protein solution 將純化之螢光蛋白EGFP溶於無菌水中,即可直接置 於進料裝置中,用於低壓氣流加速式基因投遞裝置之轟擊。 以低壓氣流加速式基因投遞裝置轟擊老鼠皮膚之實驗設計 選擇5週大之C57BL6老鼠,用剃刀除去老鼠腹部毛 髮,露出光滑之皮膚組織後’將DNA水溶液、:qNa_pei 複合物水溶液或EGFP蛋白溶液置入低壓氣流加速式基因 投遞裝置後,以低壓氣流加速式基因投遞裝置轟擊老鼠裸 露的皮膚’每隻老鼠腹部皮膚的大小約可轟擊四發,每發 的溶液體積量為20μ1。於基因搶轟擊後隔天將老鼠犧牲: 18 6 6200909579 以取下皮膚_,取下触纽_處理後可直接以榮光 顯微鏡觀察之。 以低雇氣流加速式基因投遞裝置森擊老鼠皮膚之實驗結 果 直接以DNΑ液艘溶液轟擊 分別以20、30、40與50 psi壓力的氦氣,搭配開口内 徑ltom之搶管進行低壓氣流加速式基因投遞裝置的轟 擊,每發,谷液為2〇μι,内含3jIgDNA,並於螢光顯微鏡下 観察老乳被轟擊之腹部皮膚的螢光表現情形。圖5八_圖5D 為螢光顯微鏡40倍放大下之觀察結果,依次為以2〇、3〇、 40與50 psi施打之結果。由實驗結果可看到以3〇psi與 40psi轟擊之皮膚組織有最佳之綠色螢光表現。 以PEI-DNA複合物水溶液轟擊 以30psi壓力之氦氣,搭配開口内徑1〇mm之搶管進 行低麗氣流加速式基因投遞裝置的轟擊,每發溶液為2〇μ1 之PEI-DNA複合物水溶液,内含㈣,並於螢光顯 微鏡下觀察老鼠被轟擊之腹部皮膚的螢光表現情形。如圖 6所示,當溶液中添加了 PEI之後,可使DNA劑量降低為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
Wg時’其螢光表現與每發中含3呢DNA之DNA水溶液 的結果相當之螢光。 以EGFP蛋白質轟擊 19 200909579 低壓氣加速式基因投遞裝置以氦氣% psi壓力射 擊’每發含5//gEGFP蛋白質。並在螢光顯微鏡下觀察被 A擊小鼠腹部皮膚之螢光情形。如圖7A_圖7C所示,為螢 光顯微鏡40倍放大下來觀察老鼠腹部皮膚,可觀察到 EGFP蛋白質成功且均勻地射入表皮層。 以低座氣流加速式基因投遞裝置森擊文心蘭花辮(petal 〇f Oncidium Ramsey) 以50psi壓力之氦氣,搭配開口内徑4 5mm之槍管進 行低壓氣流加速式基因投遞裝置的轟擊,每發溶液為1〇μ1 之 DNA-Spermidine-CaC12 複合物水溶液,内含 1μ§ DNA, 槍口距離目標物距離3公分,並於第二天進行GUS組織 染色’於暗室裡反應48小時後觀察GUS的呈色情形。 由前述實驗結果可見,本發明中所使用之低壓氣流加 速式基因投遞裝置並未對目標細胞組織造成嚴重之傷害。 而且本發明之低壓氣流加速式基因投遞裝置能直接將基因 DNA或其他生物性物質送入細胞内部,可以避免許多的組 織或細胞間的屏障阻礙的消耗,而成功達成基因轉殖或生 物性物質遞送之目的。 總上所述,本發明至少具有下列優點: 1.在本發明所提出的低壓氣流加速式基因投遞裝置 中’搶管結構的喷嘴具有與蓄壓艙的相同的外徑,使得槍 管結構容易缚造。 20 6 200909579 2.本發明所提出之低壓氣流加速式基因投遞裝置的連 結裝置與蓄壓艙前段連接處具有相同内徑,且漸縮部與蓄 壓艙端以圓弧相銜接,因此能消除氣體進入蓄壓艙時^ 擾流現象。 3.本發明的搶管結構的漸張部的内部輪廓是直線擴步 或疋鐘形擴張,這樣的設計可以增加加速射出的氣體並且 使射出的氣體具有高均勻度。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.本發明所提出的低壓氣流加速式基因投遞裳置具有 環狀間隔物,可使得低壓氣流加速式基因投遞裝置與欲射 擊的目標物之間保持固定間距,以降低射擊誤差。另外,' 由於本發明所提出的低壓氣流加速式基因投遞裝置環狀門 隔物具有至少一缺口,可以防止強力氣流對欲射擊 二 物產生傷害。 ^ 雖然本發明已以較佳實施例揭露如上,然其並非用以 限定本發明,任何熟習此技藝者,在不脫離本發明之精神 =範圍内,當可作些許之更動與潤飾,因此本發明之=護 範圍當視後附之申請專利範圍所界定者為準。 【圖式簡單說明】 圖1所繪示為本發明一實施例之低壓氣流加速 投遞裝置的示意圖。 土 圖2Α與圖2Β為本發明之實施例之搶管結構的示惫 圖。 ’如 圖3所繪示為圖2Α中圈示處的局部放大圖。 圖4Α所繪示為將樣本從喉部上方的進料孔投入的照 21 6 6200909579 片圖。 圖4B所繪示為從將樣本從距喉部12公分的進料孔投 入的照片圖。 圖4 C所繪示為從將樣本從距喉部2 · 5公分的進料孔投 入的照片圖。 圖4 D所繪示為從將樣本從距喉部5. 〇公分的進料孔投 入的照片圖。 圖4E所繪示為從將樣本從距喉部7·5公分的進料孔投 入的照片圖。 圖5Α為螢光顯微鏡下放大4〇倍,以20psi之氦氣轟 擊老鼠腹部皮膚所得之觀察結果 圖為螢光顯微鏡下放大40倍,以30psi之氦氣森 擊老鼠腹部皮膚所得之觀察結果 圖5C為螢光顯微鏡下放大40倍,以40psi之氦氣森 擊老鼠腹部皮膚所得之觀察結果 圖5D為榮光顯微鏡下放大40倍,以50psi之氦氣轟 擊老鼠腹部皮膚所得之觀察結果 圖6為螢光顯微鏡下放大40倍,以PEI-DNA複合物 水〉谷液’ 30psi之氦氣轟擊老鼠腹部皮膚所得之觀察結果 圖7 A所繪不為以EGFP蛋白質暴擊老鼠皮膚組織, 於螢光顯微鏡下(放大40倍)觀察所得結果(非投遞區域)。 圖7B所繪示為以EGFP蛋白質轟擊老鼠皮膚組織, 於螢光顯微鏡下(放大40倍)觀察所得結果(投遞區域i)。 圖7C所繪示為以EGFP蛋白質轟擊老鼠皮膚組織, 22 6 6200909579 於螢光顯微鏡下(放大40倍)觀察所得結果(投遞區 圖8A所緣不為以酒精(無DNA),5〇psi之氦氣爲擊文 心蘭花瓣後以GUS組織染色所得之觀察結果㈣ control) ° 圖8B所繪示為以pBI121溶液,5〇psi之氦氣轟擊文 心蘭花瓣後以GUS組織染色所得之觀察結果。 圖8C所繪示為以pRG溶液,5〇psi之氦氣轟擊文心蘭 花瓣後以GUS組織染色所得之觀察結果。 圖8D所纟會不為以pCG溶液,5〇psi之氣氣義擊文心蘭 花瓣後以GUS組織染色所得之觀察結果。 【主要元件符號說明】 100 :低壓氣流加速式基因投遞裝置 102 :氣體供應裝置 104 :搶管結構 106 :擊發裝置106 112 :氣瓶 114 :壓力調整器 116 :管路 H8 :控制閥 122 :蓄壓艙 124 =噴嘴 124a :噴嘴前端 124b :嘴嘴後端 200909579 126 :漸縮部 128 :漸張部 130 :喉部 132 :進料孔 134 :搶管外殼 136 :啟動裝置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 :漸張部128末端截面積Ae: cross-sectional area of the end of the progressive portion 128
At :喉部130截面積At : throat 130 cross-sectional area
Rt :喉部130的曲率半徑 rT :喉部130的半徑 Θ :漸張部128與中心轴之夾角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 24L 24
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TWI710724B (en) * | 2019-12-19 | 2020-11-21 | 生物鎵科技股份有限公司 | Solenoid valve, delivery device having a solenoid valve and delivery device having a solenoid valve and a disassembly-prevention safety catch |
CN111850049A (en) * | 2020-07-22 | 2020-10-30 | 庄丰如 | Low-pressure gene delivery device |
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