WO2018145495A1 - 手持式电穿孔装置 - Google Patents

手持式电穿孔装置 Download PDF

Info

Publication number
WO2018145495A1
WO2018145495A1 PCT/CN2017/110498 CN2017110498W WO2018145495A1 WO 2018145495 A1 WO2018145495 A1 WO 2018145495A1 CN 2017110498 W CN2017110498 W CN 2017110498W WO 2018145495 A1 WO2018145495 A1 WO 2018145495A1
Authority
WO
WIPO (PCT)
Prior art keywords
unit
housing
electroporation device
hand
circular hole
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2017/110498
Other languages
English (en)
French (fr)
Inventor
徐友春
程振
苏世圣
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tsinghua University
Original Assignee
Tsinghua University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Tsinghua University filed Critical Tsinghua University
Publication of WO2018145495A1 publication Critical patent/WO2018145495A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion

Definitions

  • the invention belongs to the technical field of cell transfection equipment, and in particular to a hand-held electroporation device.
  • the traditional electroporation device is bulky and inconvenient to operate.
  • the electroporation device and the electroporation cup are independent of each other, and the electroporation device has the functions of pipetting and electroporation at the same time.
  • it is first necessary to mix the cells and the foreign substances in the EP tube, and then absorb the suspension containing the cells and the foreign substances into the electroporation cup for electroporation, and then the electroporated suspension. It is then taken up into the culture dish and the operation is extremely cumbersome.
  • the present invention aims to solve at least one of the above technical problems to some extent.
  • the present invention provides a hand-held electroporation device that is convenient, fast, and efficient to operate.
  • a hand-held electroporation device includes: a housing defining a sealed receiving cavity, the sealed electrical pulse unit and a sampling unit, the housing being provided with a mounting hole, the mounting hole and The closed containment chamber is connected; the tip assembly, the tip assembly is coupled to the mounting hole, the tip assembly includes a tip, the tip defines a reservoir, and the reservoir is for storage a suspension containing cells and a foreign substance, the liquid storage chamber having a gas inlet and outlet and a liquid inlet and outlet, the sampling unit being in communication with the gas inlet and outlet, the gun head assembly being electrically connected to the electric pulse unit, the electricity
  • the electrical pulse generated by the pulse unit causes the reservoir to generate an electric field to electroporate the cells, allowing foreign matter to be introduced into the cells.
  • the cell suspension in the liquid storage chamber can be directly electroporated, compared with the conventional electric power.
  • perforating equipment it is convenient, fast and efficient.
  • hand-held electroporation device may further have the following additional technical features:
  • the handheld electroporation device further includes: a control unit, the control unit includes: a control circuit board and a display device, the control circuit board is disposed in the sealed receiving cavity and respectively The electrical pulse unit is coupled to the sampling unit, and the display device is mounted on a sidewall of the housing to be electrically connected to the control circuit board.
  • the handheld electroporation device further includes: a power supply unit, the power supply unit is disposed in the sealed receiving cavity, and the power supply unit is respectively connected to the control unit, the sampling unit, and The electrical pulse unit is electrically connected.
  • the tip assembly further includes: a mounting connector, one of the assembly connectors The end is embedded in the mounting hole, and the other end of the mounting connector protrudes from the mounting hole, and the mounting connector is provided with two electrode pins electrically connected to the electric pulse unit, the gun head and the The other end of the assembly connector is connected, and the gun head is provided with two oppositely disposed conductive plates, and the two electrode pins are respectively electrically connected to the two opposite conductive plates.
  • the tip is detachably connected to the fitting connection.
  • an insulating layer is disposed between the two conductive plates, the insulating layer defines a liquid storage cavity, and the conductive plate is attached to an outer sidewall of the insulating layer.
  • the insulating layer includes: a tubular body portion, one end of the tubular body portion is coupled to the other end of the fitting connector; a cavity portion, the cavity portion and the tubular body The other end of the portion is connected, and the cavity portion is substantially inverted triangle.
  • the hand-held electroporation device further includes: an injection plug rod assembly
  • the injection plug rod assembly includes: an injection plug rod
  • the closed receiving chamber is provided with a circular hole groove
  • the circular hole groove is in communication with the sealed receiving cavity, one end of the injection plug rod extends into the circular hole groove, and the other end of the injection plug rod is located outside the circular hole groove, the
  • the sample plug rod is movably disposed in the circular hole groove under the action of an external force; the elastic member is located in the circular hole groove, and one end of the injection plug rod is abutted against the elastic member .
  • the peripheral wall of the circular hole groove divides the sealed receiving cavity into an upper chamber and a lower chamber, and the power supply unit and the control unit are respectively disposed in the upper chamber.
  • the sampling unit and the electric pulse unit are respectively disposed in the lower chamber.
  • the housing comprises: a first housing and a second housing, the first housing and the second housing being oppositely disposed to form the sealed receiving cavity.
  • FIG. 1 is a perspective view of a hand-held electroporation device in accordance with one embodiment of the present invention
  • FIG. 2 is an exploded view of a handheld electroporation device in accordance with one embodiment of the present invention
  • Figure 3 is an exploded view of a pipette tip in accordance with one embodiment of the present invention.
  • FIG. 4 is a flow chart showing the operation of a hand-held electroporation device in accordance with one embodiment of the present invention.
  • the hand-held electroporation device 100 in accordance with an embodiment of the present invention will now be described with reference to Figures 1-3.
  • the hand-held electroporation can generally include a housing 10 and a tip assembly 40.
  • the housing 10 defines a sealed receiving cavity, and the electrical sealing unit 20 and the sampling unit 30 are sequentially disposed in the sealed receiving cavity.
  • the housing 10 is provided with a mounting hole 11, a mounting hole 11 and a sealed receiving cavity.
  • the electrical pulse unit 20 and the sampling unit 30 of the handheld electroporation device 100 are integrally disposed in the sealed receiving cavity of the housing 10. Thereby, the hand-held electroporation device 100 is simultaneously provided with the functions of sampling and electroporation.
  • the tip assembly 40 is coupled to a mounting aperture 11 that includes a tip 41 that defines a reservoir 411 for storing cells and foreign sources
  • the liquid suspension chamber 411 has a gas inlet and outlet 4111 and a liquid inlet and outlet 4112.
  • the sampling unit 30 is in communication with the gas inlet and outlet 4111.
  • the nozzle assembly 40 is electrically connected to the electrical pulse unit 20, and the electrical pulse generated by the electrical pulse unit 20 causes storage.
  • the liquid chamber 411 generates an electric field to electroporate the cells and introduce foreign substances into the cells.
  • the hand-held electroporation device 100 generally transfects the cells to include the following steps: S100 takes the cells to be transfected into the reservoir chamber 411: the sampling unit 30 suctions the reservoir chamber 411, so that the reservoir chamber 411 The gas inside is discharged from the gas inlet and outlet 4111 and forms a negative pressure, and a suspension containing cells and foreign substances is sucked into the liquid storage chamber 411 from the liquid inlet and outlet 4112 under a negative pressure; S200 electroporation is to be transfected cells: electric pulse The electrical pulse generated by unit 20 causes the reservoir chamber 411 to generate an electric field to electroporate the cells and introduce foreign species into the cells; S300 transfers the electroporated cell suspension: through the sampling unit 30 to the reservoir chamber 411 Air is introduced, and gas enters the liquid storage chamber 411 from the gas inlet and outlet 4111 so that the cell suspension is discharged from the liquid inlet and outlet 4112, and is transferred to a culture dish for cultivation.
  • S100 takes the cells to be transfected into the reservoir
  • the parameters related to the electrical pulse unit 20 are preset before sampling: for example, the type of voltage pulse, the voltage amplitude, the width of each pulse, the number of consecutive pulses, the interval between two pulses, and the sampling unit is preset. Injection volume of 30.
  • the hand-held electroporation device 100 of the embodiment of the present invention by directly arranging the electric pulse unit 20 and the sampling unit 30 in the sealed accommodating cavity, the cell suspension in the liquid storage chamber 411 can be directly electroporated. It is easy to use, fast and efficient compared to traditional electroporation equipment.
  • the housing 10 includes a first housing 13 and a second housing 14, and the first housing 13 and the second housing 14 are oppositely disposed. Out of the containment chamber.
  • the first housing 13 and the second housing 14 are detachably connected, for example, the first housing 13 and the second housing 14 may be connected by screws or snaps.
  • the handheld electroporation device 100 further includes: a control unit 50.
  • the control unit 50 includes a control circuit board 51 and a display device 52.
  • the control circuit board 51 is disposed in the sealed housing chamber and is respectively connected to the electric pulse unit 20 and the sampling unit 30, and the display device 52 is mounted on the side wall of the housing 10 to be electrically connected to the control circuit board 51.
  • the type of voltage pulse of the electric pulse unit 20 the voltage amplitude, the width of each pulse, and the continuous pulse
  • the number and the interval between the two pulses can be controlled by the control circuit board 51, and the sampling volume of the sampling unit 30 can also be controlled by the control circuit board 51.
  • all of the set parameters can be displayed by the display device 52. Thereby, the operator is further facilitated to set different parameters for different cell types to meet the needs of different experimental requirements, thereby improving the versatility of the handheld electroporation device 100.
  • the housing 10 can be provided with an adjustment knob 21 for adjusting the parameters associated with the electrical pulse unit 20 and a button 31 for performing the sampling operation of the sampling unit 30, thereby further improving the ease of operation of the handheld electroporation device 100.
  • the handheld electroporation device 100 may further include: a power supply unit 60.
  • the power supply unit 60 is disposed in the sealed housing chamber, and the power supply unit 60 is electrically connected to the control unit 50, the sampling unit 30, and the electrical pulse unit 20, respectively. That is, the power supply unit 60 can supply power to the control unit 50, the sampling unit 30, and the electrical pulse unit 20 at the same time.
  • the design of the entire handheld electroporation device 100 can be made more reasonable, and the need for the operator to use the hand-held electroporation device 100 in different occasions can be satisfied.
  • the tip assembly 40 further includes an assembly connector 42.
  • One end of the assembly connecting body 42 is embedded in the mounting hole 11, and the other end of the mounting connecting body 42 protrudes from the mounting hole 11, and the mounting connecting body 42 is provided with two electrode pins 43 electrically connected to the electric pulse unit 20, the gun head 41 and the assembly
  • the other end of the connecting body 42 is connected, and the tip 41 is provided with two opposite conductive plates 412, and the two electrode pins 43 are electrically connected to the two conductive plates 412, respectively.
  • one end of the two electrode pins 43 is electrically connected to the two conductive plates 412, and the other end of the two electrode pins 43 is electrically connected to the electric pulse unit 20.
  • the pulse electric field generated by the electric pulse unit 20 can pass through the two electrode pins 43. It is transferred to the conductive plate 412 to form a pulsed electric field between the two oppositely disposed conductive plates 412, so that cells in the reservoir chamber 411 can be electroporated.
  • the two electrode pins 43 may be respectively connected to the outer sidewalls of the two conductive plates 412, and the distance d 1 between the two electrode pins 43 is substantially the distance d 2 between the two conductive plates 412.
  • the spacing d 1 of the two electrode pins 43 may be between 0.2 and 2 mm.
  • d 1 is 1 mm.
  • the tip 41 is detachably coupled to the fitting connector 42.
  • the tip 41 can be removed from the assembly connector 42 for cleaning, or a disposable tip 41 can be used, and a new tip 41 can be replaced after each electroporation is completed. Thereby, cross-contamination during cell transfection can be avoided.
  • an insulating layer 413 is disposed between the two conductive plates 412.
  • the insulating layer 413 defines a liquid storage chamber 411, and the conductive plate 412 is attached to the insulating layer.
  • the insulating layer 413 is sandwiched between the two conductive plates 412 such that a stable pulsed electric field can be formed between the two conductive plates 412 to effectively electroporate the cells in the reservoir chamber 411.
  • the insulating layer 413 includes a tube portion 4131 and a cavity portion 4132, and one end of the tube portion 4131 is connected to the other end of the mounting connector 42.
  • the cavity portion 4132 is coupled to the other end of the tubular body portion 4131, and the cavity portion 4132 is generally an inverted triangular shape.
  • the two conductive plates 412 and the insulating layer 413 have a shape and structure, and the two conductive plates 412 are coated on the outer side of the insulating layer 413.
  • an adhesive layer may be disposed between the insulating layer 413 and the conductive plate 412, and the adhesive layer is adhered The junction strengthens the connection strength between the insulating layer 413 and the conductive plate 412.
  • the handheld electroporation device 100 can further include an injection plunger assembly 70.
  • the plunger rod assembly 70 includes an injection stopper rod 71 and an elastic member 72.
  • the closed receiving cavity is provided with a circular hole groove 12, and the circular hole groove 12 communicates with the sealed receiving cavity.
  • One end of the injection plug rod 71 extends into the circular hole groove 12, and the other end of the injection plug 71 is injected.
  • the injection plug rod 71 is movably disposed in the circular hole groove 12 by an external force.
  • the elastic member 72 is located in the circular hole groove 12, and one end of the injection stopper rod 71 abuts against the elastic member 72.
  • the mounting through hole of the handheld electroporation device 100 may be provided with a pipette, and the handheld electroporation device 100 has the function of a common pipetting gun.
  • the injection stopper rod 71 By pressing the injection stopper rod 71, the injection stopper rod 71 is moved from the other end (the outer end of the injection stopper rod 71) toward one end (the inner end of the injection stopper rod 71), and the elastic member 72 is deformed after being subjected to the force.
  • the gas in the circular orifice 12 flows into the sealed accommodating chamber, forcing the gas in the pipette to be discharged.
  • the restoring force of the elastic member 72 drives the injection stopper 71 from one end to the other.
  • the movement in one end direction causes the air pressure of the sealed accommodating chamber to decrease, the liquid is sucked into the pipette, and when the injection stopper rod 71 is pressed again, the liquid in the pipette is discharged, thereby causing the hand-held electroporation device 100 to complete the pipetting. And the process of draining.
  • the peripheral wall of the circular hole groove 12 can divide the sealed receiving cavity into an upper chamber and a lower chamber, the power supply unit 60 and the control unit 50 are disposed in the upper chamber, and the sampling unit and the electric pulse unit 20 are disposed in the lower chamber.
  • the terms “installation”, “connected”, “connected”, “fixed” and the like shall be understood broadly, and may be either a fixed connection or a detachable connection, unless explicitly stated and defined otherwise. , or connected integrally; may be mechanical connection or electrical connection; may be directly connected, or may be indirectly connected through an intermediate medium, and may be internal communication between the two elements.
  • installation shall be understood broadly, and may be either a fixed connection or a detachable connection, unless explicitly stated and defined otherwise.
  • , or connected integrally may be mechanical connection or electrical connection; may be directly connected, or may be indirectly connected through an intermediate medium, and may be internal communication between the two elements.
  • the specific meanings of the above terms in the present invention can be understood on a case-by-case basis.
  • the first feature "on” or “under” the second feature may include direct contact of the first and second features, and may also include first and second features, unless otherwise specifically defined and defined. It is not in direct contact but through additional features between them.
  • the first feature “above”, “above” and “above” the second feature includes the first feature directly above and above the second feature, or merely indicating that the first feature level is higher than the second feature.
  • the first feature “below”, “below” and “below” the second feature includes the first feature directly above and above the second feature, or merely the first feature level being less than the second feature.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Sustainable Development (AREA)
  • Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Electromagnetism (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

一种手持式电穿孔装置（100），包括：壳体（10），壳体（10）限定出密闭容纳腔，密闭容纳腔内依次设有电脉冲单元（20）和取样单元（30），壳体（10）设有安装孔（11），安装孔（11）与密闭容纳腔相通；枪头组件（40），枪头组件（40）与安装孔（11）连接，枪头组件（40）包括枪头（41），枪头（41）限定出储液腔（411），储液腔（411）用于储存含有细胞和外源物质的悬浮液，储液腔（411）具有气体出入口（4111）和液体出入口（4112），取样单元（30）与气体出入口（4111）连通，枪头组件（40）与电脉冲单元（20）电连接，电脉冲单元（20）所产生的电脉冲使得储液腔内产生电场以对细胞进行电穿孔，并使得外源物质导入至细胞内。

Description

手持式电穿孔装置 技术领域
本发明属于细胞转染设备技术领域，具体地，涉及一种手持式电穿孔装置。
背景技术
传统的电穿孔设备体积庞大，操作不便，电穿孔设备与电穿孔杯相互独立，电穿孔设备不同时具备移液和电穿孔的功能。在电穿孔实验中，首先需要在EP管中混合细胞和外源物质之后，再将含有细胞和外源物质的悬浮液吸取到电穿孔杯内进行电穿孔，接着将经过电穿孔后的悬浮液再吸取到培养皿中，操作极其繁琐。
发明内容
本发明旨在至少在一定程度上解决上述技术问题之一。
为此，本发明提出一种手持式电穿孔装置，该手持式电穿孔装置操作方便且快捷高效。
根据本发明实施例的手持式电穿孔装置包括：壳体，所述壳体限定出密闭容纳腔，所述密闭电脉冲单元和取样单元，所述壳体设有安装孔，所述安装孔与所述密闭容纳腔相连；枪头组件，所述枪头组件与所述安装孔连接，所述枪头组件包括枪头，所述枪头限定出储液腔，所述储液腔用于储存含有细胞和外源物质的悬浮液，所述储液腔具有气体出入口和液体出入口，所述取样单元与所述气体出入口连通，所述枪头组件与所述电脉冲单元电连接，所述电脉冲单元所产生的电脉冲使得储液腔产生电场对细胞进行电穿孔，使得外源物质导入至细胞内。
根据本发明实施例的手持式电穿孔装置，通过将电脉冲单元和取样单元集成设置于密闭容纳腔内，从而可以实现对储液腔内的细胞悬浮液直接进行电穿孔，相比于传统电穿孔设备而言，使用方便且快捷高效。
另外，根据本发明实施例的手持式电穿孔装置，还可以具有如下附加的技术特征：
根据本发明的一个实施例，所述手持式电穿孔装置还包括：控制单元，所述控制单元包括：控制电路板和显示装置，所述控制电路板设置于所述密闭容纳腔内且分别与所述电脉冲单元和所述取样单元连接，所述显示装置安装在所述壳体的侧壁上与所述控制电路板电连接。
根据本发明的一个实施例，所述手持式电穿孔装置还包括：供电单元，所述供电单元设在所述密闭容纳腔内，所述供电单元分别与所述控制单元、所述取样单元和所述电脉冲单元电连接。
根据本发明的一个实施例，所述枪头组件还包括：装配连接体，所述装配连接体的一 端嵌入所述安装孔内，所述装配连接体的另一端伸出所述安装孔，所述装配连接体设有与所述电脉冲单元电连接的两个电极针，所述枪头与所述装配连接体的另一端连接，所述枪头设有两个相对设置的导电板，两个所述电极针分别与两个所述相对导电板电连接。
根据本发明的一个实施例，所述枪头与所述装配连接体可拆卸地连接。
根据本发明的一个实施例，两个所述导电板之间设有绝缘层，所述绝缘层限定出储液腔，所述导电板贴合在所述绝缘层的外侧壁。
根据本发明的一个实施例，所述绝缘层包括：管体部，所述管体部的一端与所述装配连接体的另一端连接；腔体部，所述腔体部与所述管体部的另一端连接，所述腔体部大体为倒三角型。
根据本发明的一个实施例，所述手持式电穿孔装置还包括：进样塞杆组件，所述进样塞杆组件包括：进样塞杆，所述密闭容纳腔内设有圆孔槽，所述圆孔槽与所述密闭容纳腔连通，所述进样塞杆的一端伸入所述圆孔槽内，所述进样塞杆的另一端位于所述圆孔槽外，所述进样塞杆在外力的作用下可移动地设在所述圆孔槽内；弹性件，所述弹性件位于所述圆孔槽内，所述进样塞杆的一端止抵于所述弹性件。
根据本发明的一个实施例，所述圆孔槽的周壁将所述密闭容纳腔分隔成上腔室和下腔室，所述供电单元和所述控制单元分别设在所述上腔室内，所述取样单元和电脉冲单元分别设在所述下腔室。
根据本发明的一个实施例，所述壳体包括：第一壳体和第二壳体，所述第一壳体和所述第二壳体相对设置构设出所述密闭容纳腔。
本发明的附加方面和优点将在下面的描述中部分给出，部分将从下面的描述中变得明显，或通过本发明的实践了解到。
附图说明
本发明的上述和/或附加的方面和优点从结合下面附图对实施例的描述中将变得明显和容易理解，其中：
图1是根据本发明一个实施例的手持式电穿孔装置的立体图；
图2是根据本发明一个实施例的手持式电穿孔装置的分解图；
图3是根据本发明一个实施例的枪头的分解图；
图4是根据本发明一个实施例的手持式电穿孔装置的工作流程图。
具体实施方式
下面详细描述本发明的实施例，所述实施例的示例在附图中示出，其中自始至终相同或类似的标号表示相同或类似的元件或具有相同或类似功能的元件。下面通过参考附图描 述的实施例是示例性的，旨在用于解释本发明，而不能理解为对本发明的限制。
下面参照图1-图3描述根据本发明实施例的手持式电穿孔装置100。该手持式电穿孔大体可以包括：壳体10和枪头组件40。
具体地，如图2所示，壳体10限定出密闭容纳腔，密闭容纳腔内依次设有电脉冲单元20和取样单元30，壳体10设有安装孔11，安装孔11与密闭容纳腔相通。也就是说，该手持式电穿孔装置100的电脉冲单元20和取样单元30均集成设置于壳体10的密闭容纳腔内。由此，使得手持式电穿孔装置100同时具备取样和电穿孔的功能。
参照图1结合图3所示，枪头组件40与安装孔11连接，枪头组件40包括枪头41，枪头41限定出储液腔411，储液腔411用于储存含有细胞和外源物质的悬浮液，储液腔411具有气体出入口4111和液体出入口4112，取样单元30与气体出入口4111连通，枪头组件40与电脉冲单元20电连接，电脉冲单元20所产生的电脉冲使得储液腔411产生电场以对细胞进行电穿孔，并使得外源物质导入至细胞内。
参照图4所示，手持式电穿孔装置100对细胞转染大体包括如下步骤：S100取待转染细胞至储液腔411：取样单元30对储液腔411进行抽吸，使得储液腔411内的气体从气体出入口4111排出并形成负压，含有细胞和外源物质的悬浮液从液体出入口4112在负压作用下被吸入至储液腔411中；S200电穿孔待转染细胞：电脉冲单元20所产生的电脉冲使得储液腔411产生电场以对细胞进行电穿孔，并使得外源物质导入至细胞内；S300转移经过电穿孔的细胞悬浮液：通过取样单元30对储液腔411通入空气，气体从气体出入口4111进入储液腔411内使得细胞悬浮液从液体出入口4112排出，并被转移至培养皿中进行培养。
可以理解的是，在取样之前预先设定电脉冲单元20相关参数：例如电压脉冲的类型、电压幅度、每个脉冲的宽度、连续脉冲数目、两个脉冲的间隔，同时，预先设定取样单元30的进样体积。
根据本发明实施例的手持式电穿孔装置100，通过将电脉冲单元20和取样单元30集成设置于密闭容纳腔内，从而可以实现对储液腔411内的细胞悬浮液直接进行电穿孔，相比于传统电穿孔设备而言，使用方便且快捷高效。
在本发明的一些实施例中，如图1和图2所示，壳体10包括：第一壳体13和第二壳体14，第一壳体13和第二壳体14相对设置构设出密闭容纳腔。其中，第一壳体13和第二壳体14可拆卸地连接，例如，可以第一壳体13和第二壳体14可以通过螺钉或卡扣方式连接。由此，简化手持式电穿孔装置100的制造工序，降低生产成本。
可选地，手持式电穿孔装置100还包括：控制单元50。控制单元50包括：控制电路板51和显示装置52。控制电路板51设置于密闭容纳腔内且分别与电脉冲单元20和取样单元30连接，显示装置52安装在壳体10的侧壁上与控制电路板51电连接。
也就是说，电脉冲单元20的电压脉冲的类型、电压幅度、每个脉冲的宽度、连续脉冲 数目以及两个脉冲的间隔可以通过控制电路板51进行控制，取样单元30的取样体积也可以通过控制电路板51进行控制。且所有设定的参数可以通过显示装置52显示出来。由此，进一步便于操作者针对不同细胞种类设定不同参数，以满足不同实验要求的需要，从而提高手持式电穿孔装置100的通用性。
有利地，壳体10上可以设有调节电脉冲单元20相关参数的调节旋钮21和用于执行取样单元30取样动作的按钮31，由此，进一步提高手持式电穿孔装置100操作的便利性。
可选地，手持式电穿孔装置100还可以包括：供电单元60。供电单元60设在密闭容纳腔内，供电单元60分别与控制单元50、取样单元30和电脉冲单元20电连接。也就是说，供电单元60可以同时向控制单元50、取样单元30和电脉冲单元20提供电源。由此，可以使得整个手持式电穿孔装置100的设计更加合理，满足操作者在不同场合使用手持式电穿孔装置100的需要。
在本发明另一些实施例中，如图1结合图2所示，枪头组件40还包括：装配连接体42。装配连接体42的一端嵌入安装孔11内，装配连接体42的另一端伸出安装孔11，装配连接体42设有与电脉冲单元20电连接的两个电极针43，枪头41与装配连接体42的另一端连接，枪头41设有两个相对设置的导电板412，两个电极针43分别与两个导电板412电连接。换言之，两个电极针43的一端与两个导电板412电连接，两个电极针43的另一端与电脉冲单元电20连接，电脉冲单元20所产生的脉冲电场可以通过两个电极针43传递至导电板412，从而在两个相对设置的导电板412之间形成脉冲电场，进而可以对储液腔411内的细胞进行电穿孔。
可选地，两个电极针43可以分别与两个导电板412的外侧壁连接，两个电极针43之间的间距d1大体为两个导电板412之间的距离d2。
可选地，两个电极针43的间距d1可以为0.2-2毫米之间。优选地，d1为1毫米。
一些实施例中，枪头41与装配连接体42可拆卸地连接。这样，在完成一次电穿孔之后，可以将枪头41从装配连接体42取下来清洗，或者使用一次性枪头41，每次电穿孔结束后更换使用新的枪头41。由此，可以避免细胞转染过程中出现交叉污染。
在本发明的一个具体实施例中，参照图2结合图3所示，两个导电板412之间设有绝缘层413，绝缘层413限定出储液腔411，导电板412贴合在绝缘层413的外侧壁。换言之，绝缘层413夹设在两个导电板412之间，从而使得两个导电板412之间可以形成稳定的脉冲电场，有效对储液腔411内的细胞进行电穿孔。
进一步地，绝缘层413包括：管体部4131和腔体部4132，管体部4131的一端与装配连接体42的另一端连接。腔体部4132与管体部4131的另一端连接，腔体部4132大体为倒三角型。如图3所示，两个导电板412和绝缘层413的形状结构相适配，两个导电板412包覆在绝缘层413的外侧。其中，绝缘层413与导电板412之间可以设有粘结层，通过粘 结层加强绝缘层413与导电板412之间的连接强度。
在本发明再一些实施例中，手持式电穿孔装置100还可以包括：进样塞杆组件70。进样塞杆组件70包括：进样塞杆71和弹性件72。如图2所示，密闭容纳腔内设有圆孔槽12，圆孔槽12与密闭容纳腔连通，进样塞杆71的一端伸入圆孔槽12内，进样塞杆71的另一端位于圆孔槽12外，进样塞杆71在外力的作用下可移动地设在圆孔槽12内。弹性件72位于圆孔槽12内，进样塞杆71的一端止抵于弹性件72。
可选地，手持式电穿孔装置100的安装通孔可以设有移液管，手持式电穿孔装置100具有普通移液枪的功能。通过按压进样塞杆71使得进样塞杆71从另一端(进样塞杆71的外端)向一端(进样塞杆71的内端)方向移动，弹性件72受力之后发生形变，圆孔槽12内气体向密闭容纳腔流动，迫使移液管内的气体被排出，当进样塞杆71所受的外力消失之后，弹性件72的回复力驱动进样塞杆71从一端向另一端方向移动使得密闭容纳腔的气压减小，液体被吸进移液管中，当再次按压进样塞杆71时，移液管内的液体被排出，从而使得手持式电穿孔装置100完成吸液和排液的过程。
其中，圆孔槽12的周壁可以将密闭容纳腔分隔成上腔室和下腔室，供电单元60和控制单元50设在上腔室内，取样单元和电脉冲单元20设在下腔室。由此，可以提高手持式电穿孔装置100结构设计的合理性，保证密闭容纳腔的密封性，提高进样精度。
在本发明的描述中，需要理解的是，术语“上”、“下”“、底”、“内”、“外”等指示的方位或位置关系为基于附图所示的方位或位置关系，仅是为了便于描述本发明和简化描述，而不是指示或暗示所指的装置或元件必须具有特定的方位、以特定的方位构造和操作，因此不能理解为对本发明的限制。
在本发明中，除非另有明确的规定和限定，术语“安装”、“相连”、“连接”、“固定”等术语应做广义理解，例如，可以是固定连接，也可以是可拆卸连接，或一体地连接；可以是机械连接，也可以是电连接；可以是直接相连，也可以通过中间媒介间接相连，可以是两个元件内部的连通。对于本领域的普通技术人员而言，可以根据具体情况理解上述术语在本发明中的具体含义。
在本发明中，除非另有明确的规定和限定，第一特征在第二特征之“上”或之“下”可以包括第一和第二特征直接接触，也可以包括第一和第二特征不是直接接触而是通过它们之间的另外的特征接触。而且，第一特征在第二特征“之上”、“上方”和“上面”包括第一特征在第二特征正上方和斜上方，或仅仅表示第一特征水平高度高于第二特征。第一特征在第二特征“之下”、“下方”和“下面”包括第一特征在第二特征正上方和斜上方，或仅仅表示第一特征水平高度小于第二特征。
在本说明书的描述中，参考术语“一个实施例”、“一些实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点 包含于本发明的至少一个实施例或示例中。在本说明书中，对上述术语的示意性表述不一定指的是相同的实施例或示例。而且，描述的具体特征、结构、材料或者特点可以在任何的一个或多个实施例或示例中以合适的方式结合。
尽管上面已经示出和描述了本发明的实施例，可以理解的是，上述实施例是示例性的，不能理解为对本发明的限制，本领域的普通技术人员在不脱离本发明的原理和宗旨的情况下在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。

Claims (10)

  1. 一种手持式电穿孔装置，其特征在于，包括：
    壳体，所述壳体限定出密闭容纳腔，所述密闭容纳腔内依次设有电脉冲单元和取样单元，所述壳体设有安装孔，所述安装孔与所述密闭容纳腔相通；
    枪头组件，所述枪头组件与所述安装孔连接，所述枪头组件包括枪头，所述枪头限定出储液腔，所述储液腔用于储存含有细胞和外源物质的悬浮液，所述储液腔具有气体出入口和液体出入口，所述取样单元与所述气体出入口连通，所述枪头组件与所述电脉冲单元电连接，所述电脉冲单元所产生的电脉冲使得储液腔内产生电场以对细胞进行电穿孔，并使得外源物质导入至细胞内。
  2. 根据权利要求1所述的手持式电穿孔装置，其特征在于，还包括：控制单元，所述控制单元包括：控制电路板和显示装置，所述控制电路板设置于所述密闭容纳腔内且分别与所述电脉冲单元和所述取样单元连接，所述显示装置安装在所述壳体的侧壁上与所述控制电路板电连接。
  3. 根据权利要求2所述的手持式电穿孔装置，其特征在于，还包括：供电单元，所述供电单元设在所述密闭容纳腔内，所述供电单元分别与所述控制单元、所述取样单元和所述电脉冲单元电连接。
  4. 根据权利要求1所述的手持式电穿孔装置，其特征在于，所述枪头组件还包括：
    装配连接体，所述装配连接体的一端嵌入所述安装孔内，所述装配连接体的另一端伸出所述安装孔，所述装配连接体设有与所述电脉冲单元电连接的两个电极针，所述枪头与所述装配连接体的另一端连接，所述枪头设有两个相对设置的导电板，两个所述电极针分别与两个所述导电板电连接。
  5. 根据权利要求4所述的手持式电穿孔装置，其特征在于，所述枪头与所述装配连接体可拆卸地连接。
  6. 根据权利要求4所述的手持式电穿孔装置，其特征在于，两个所述导电板之间设有绝缘层，所述绝缘层限定出储液腔，所述导电板贴合在所述绝缘层的外侧壁。
  7. 根据权利要求6所述的手持式电穿孔装置，其特征在于，所述绝缘层包括：
    管体部，所述管体部的一端与所述装配连接体的另一端连接；
    腔体部，所述腔体部与所述管体部的另一端连接，所述腔体部大体为倒三角型。
  8. 根据权利要求4所述的手持式电穿孔装置，其特征在于，还包括：进样塞杆组件，所述进样塞杆组件包括：
    进样塞杆，所述密闭容纳腔内设有圆孔槽，所述圆孔槽与所述密闭容纳腔连通，所述进样塞杆的一端伸入所述圆孔槽内，所述进样塞杆的另一端位于所述圆孔槽外，所述进样 塞杆在外力的作用下可移动地设在所述圆孔槽内；
    弹性件，所述弹性件位于所述圆孔槽内，所述进样塞杆的一端止抵于所述弹性件。
  9. 根据权利要求8所述的手持式电穿孔装置，其特征在于，所述圆孔槽的周壁将所述密闭容纳腔分隔成上腔室和下腔室，所述供电单元和所述控制单元设分别在所述上腔室内，所述取样单元和电脉冲单元分别设在所述下腔室。
  10. 根据权利要求1所述的手持式电穿孔装置，其特征在于，所述壳体包括：第一壳体和第二壳体，所述第一壳体和所述第二壳体相对设置构设出所述密闭容纳腔。
PCT/CN2017/110498 2017-02-09 2017-11-10 手持式电穿孔装置 Ceased WO2018145495A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201710071412.8 2017-02-09
CN201710071412.8A CN107058095B (zh) 2017-02-09 2017-02-09 手持式电穿孔装置

Publications (1)

Publication Number Publication Date
WO2018145495A1 true WO2018145495A1 (zh) 2018-08-16

Family

ID=59599239

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/110498 Ceased WO2018145495A1 (zh) 2017-02-09 2017-11-10 手持式电穿孔装置

Country Status (2)

Country Link
CN (1) CN107058095B (zh)
WO (1) WO2018145495A1 (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107058095B (zh) * 2017-02-09 2020-07-28 清华大学 手持式电穿孔装置

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5318514A (en) * 1992-08-17 1994-06-07 Btx, Inc. Applicator for the electroporation of drugs and genes into surface cells
US20060281182A1 (en) * 2004-04-19 2006-12-14 Invitrogen Corporation Electroporation apparatus and methods
CN103060191A (zh) * 2004-06-12 2013-04-24 英杰生命新加坡私人有限公司 一种电穿孔系统
WO2013166203A2 (en) * 2012-05-01 2013-11-07 Life Technologies Corporation Pippette devices, systems, and methods of use thereof
CN103937669A (zh) * 2014-03-18 2014-07-23 苏州文曲生物微系统有限公司 一种手持电穿孔装置
CN105143436A (zh) * 2013-02-20 2015-12-09 陈剑 电穿孔方法与设备
CN107058095A (zh) * 2017-02-09 2017-08-18 清华大学 手持式电穿孔装置

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7771984B2 (en) * 2004-05-12 2010-08-10 Maxcyte, Inc. Methods and devices related to a regulated flow electroporation chamber
KR101084528B1 (ko) * 2008-04-15 2011-11-18 인비트로겐 싱가포르 피티이. 엘티디. 전기천공 장치용 파이펫 팁

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5318514A (en) * 1992-08-17 1994-06-07 Btx, Inc. Applicator for the electroporation of drugs and genes into surface cells
US20060281182A1 (en) * 2004-04-19 2006-12-14 Invitrogen Corporation Electroporation apparatus and methods
CN103060191A (zh) * 2004-06-12 2013-04-24 英杰生命新加坡私人有限公司 一种电穿孔系统
WO2013166203A2 (en) * 2012-05-01 2013-11-07 Life Technologies Corporation Pippette devices, systems, and methods of use thereof
CN105143436A (zh) * 2013-02-20 2015-12-09 陈剑 电穿孔方法与设备
CN103937669A (zh) * 2014-03-18 2014-07-23 苏州文曲生物微系统有限公司 一种手持电穿孔装置
CN107058095A (zh) * 2017-02-09 2017-08-18 清华大学 手持式电穿孔装置

Also Published As

Publication number Publication date
CN107058095B (zh) 2020-07-28
CN107058095A (zh) 2017-08-18

Similar Documents

Publication Publication Date Title
KR101084528B1 (ko) 전기천공 장치용 파이펫 팁
EP3593656A1 (en) Ultrasonic electronic cigarette atomizer
EP3977878A1 (en) E-liquid injection method for cartridge, cartridge and electronic cigarette
RU2006143560A (ru) Устройство для электропорации с удлиненным полым элементом
CN113495087B (zh) Poct血细胞分析仪及其检测方法
CN107091875B (zh) 一种可自动处理样品的手持吸引式电化学检测装置
WO2020248400A1 (zh) 雾化组件及雾化装置
CN110169602A (zh) 电子雾化装置及其雾化器和用于雾化器的雾化壳体
WO2021185319A1 (zh) 雾化器及气溶胶发生装置、注液系统、注液方法
WO2018145495A1 (zh) 手持式电穿孔装置
CN207428419U (zh) 雾化装置及其电子烟
CN205127992U (zh) 一种电动移液器
CN207428418U (zh) 雾化装置及其电子烟
CN207042376U (zh) 超声波震动细胞混匀移液枪
CN108642089B (zh) 超声微泡介导转染用装置
CN210581535U (zh) 一种可自动加液的电动刷
EP4678032A1 (en) Atomizer and electronic atomization device comprising same
CN115300784B (zh) 一种用于物质递送的微流控芯片
CN210017899U (zh) 雾化器及电子烟
KR101447899B1 (ko) 전기천공 장치용 파이펫 팁
CN213490873U (zh) 一种香薰液容纳仓、雾化装置及雾化器
CN211374411U (zh) 一种石油产品密度测定器
CN210058297U (zh) 移液器吸头组件
KR20100118098A (ko) 전기천공 장치용 파이펫 팁
KR101492599B1 (ko) 전기천공 장치용 파이펫 팁

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17896028

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17896028

Country of ref document: EP

Kind code of ref document: A1