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WO2008051169A1 - Chambre d'electrodes de pointe pour electroporation de petit volume - Google Patents

Chambre d'electrodes de pointe pour electroporation de petit volume Download PDF

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Publication number
WO2008051169A1
WO2008051169A1 PCT/SI2007/000036 SI2007000036W WO2008051169A1 WO 2008051169 A1 WO2008051169 A1 WO 2008051169A1 SI 2007000036 W SI2007000036 W SI 2007000036W WO 2008051169 A1 WO2008051169 A1 WO 2008051169A1
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WO
WIPO (PCT)
Prior art keywords
electrodes
housing
tip electrode
electrode chamber
cells
Prior art date
Application number
PCT/SI2007/000036
Other languages
English (en)
Inventor
Katja Trontelj
Matej Rebersek
Damjan Miklavcic
Original Assignee
Univerza V Ljubljani, Fakulteta Za Elektrotehniko
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 Univerza V Ljubljani, Fakulteta Za Elektrotehniko filed Critical Univerza V Ljubljani, Fakulteta Za Elektrotehniko
Publication of WO2008051169A1 publication Critical patent/WO2008051169A1/fr

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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

  • Subject of present invention is an apparatus for electroporation with pipetter, multi-electrodes design with minimized inhomogeneous electric field and solid filler for electrode chambers to minimize inhomogeneous electric field.
  • the present invention relates to the tip electrodes for small volume electroporation, electrofusion and gene transfection.
  • the invention resides in the field of electroporation of biological cells in suspension and more precisely in the field of techniques and devices for electroporation with the aim of cell membrane electropermeabilization and consequently electro insertion of any active pharmaceutical N chemical or biological active substances, gene transfection and cell fusion.
  • the issue of the invention are the electrodes for quick and efficient electroporation of small and controlled volumes of cell suspensions, which retain their high viability because of minimal mechanical manipulation and the possibility of application of electric pulses in different directions thus increasing the membrane area of cell being electroporated and therefore efficiency of electroporation without a significant loss in cell survival.
  • Electroporation also termed as electropermeabilization
  • electropermeabilization Electroporation of cells is used for different purposes that employ the permeabilized state of the cell membrane.
  • Electro insertion or extraction represents introducing or extracting different molecules like drugs, proteins or genes into the cells.
  • the insertion of small molecules is efficiently achieved by application of several short rectangular high voltage pulses. This is now efficiently used in electrochemotherapy for introducing drugs into the tumor cells of the patients in situ.
  • Electroextraction is used for extracting products from yeast in downstream processes.
  • Gene electrotransfection in vitro and in vivo represents the insertion of plasmids containing appropriate genetic material into cells.
  • DNA is a large molecule and was found to be efficiently introduced with the use of exponentially decaying pulses or with the combination of short high voltage and longer low voltage pulses.
  • Another related procedure is electrofusion of cells. It consists of two steps. One is making cell membranes fusogene by means of electroporation and another is achieving close contacts between the cells. It can be achieved either before or very soon (in first couple of minutes) after the electroporation until membranes are still in the fusogene state.
  • Electroporated cells have damaged membranes and are substantially more sensitive to pipetting that exerts shear forces to the cells. This appears as a problem for example at the electrofusion protocols, where contacts between cells are achieved after the electroporation. Cells have to be moved from chamber to the centrifuge tube with micropipette immediately after the electroporation. Small volume
  • Patent US6746441 describes a flow through chamber apparatus with rotating electric fields produced by more than two (four or six) electrodes. Yet this is a flow through system for ex vivo gene therapy and does not allow for small volume operation.
  • Patent US2005048651 describes the tip electrodes that also include any different kinds of conductive surface N electrodes. It is designed for the delivery of different substances from inside the tip to the targets (cells, etc.) that are outside of the tip synchronized with application of the electric pulses. It does not foresee however cells to be inserted into the lumen of the tip electrodes and electrically manipulated there.
  • a significant problem is the process of fusion of two different types of cells. This is the case for example in hybridoma technology for monoclonal antibodies production where lymphocytes B are to be fused with myeloma fusion partners. The later can be almost twice as large as lymphocytes. Common electrical treatment of both cell types is not optimal in this case. Separate electroporation and contacting cells after the electroporation is though necessary for optimal result of fusion process. The same problem exists with any pair of cells that are differently sensitive to electric field for any other reason but different sizes.
  • Cuvette as described in WO03/050232 includes the inserts of support structure that holds a porous membrane and facilitates membrane-based fusion of cells.
  • both types of cells are electroporated at the same time. In that case, either cells with smaller size are sub optimally permeabilized or larger cells are damaged.
  • the systems described in DE10359189 and DE10359190 are described that avoid mechanical manipulation since cells are treated in the culture dishes, though for subsequent fusion of separately electroporated cells transferring of cells still has to be performed by pipetting.
  • Electric field that is applied (with defined electric pulses) is not homogeneous in the whole space between the electrodes but only in the middle part of that space. Only the cells that are in that middle part are exposed to desired electric field.
  • the problem not yet solved is electroporation of predefined small volumes of cells in suspension. This will reduce costs or enable gene transfection, protein insertion and cell fusion experiments when the quantity of the proteins, plasmids or the cells is limited.
  • the object of this invention is an effective electroporation of cells with the pulses delivered to cells in different directions is also to be enabled at the same time with the possibility of fast manipulation of cells with minimal mechanical stress.
  • Another object of the present invention is to provide an apparatus for electroporation mediated, in vitro, cell fusion. It is a further object of the present invention to provide a method for fusion of two different types of cells, mediated by distinct electroporation with different values of electrical parameters.
  • the present invention provides also an interior of the tip electrode chamber made of material with the conductance that is comparable to the conductance of the medium of the cell suspension. This assures the retention of cells in the central area of the chamber and the achievement of a desired homogeneous electric field in that area.
  • the present invention allows cells to be confined to that middle part of the lumen by incorporating for example a semi conductive material or composite material of the conductance that is comparable to the conductance of the medium of the cell suspension in the lumen of the tip electrode chamber. It provides the homogeneity of the field because of the conductance that is similar to the conductance of the medium of the cell suspension. Accordingly, it is the primary object of the present invention to provide tip electrode chamber for electroporation of small and controlled volumes of cells with minimal mechanical manipulation and consecutively minimal mechanical stress.
  • the described invention allows for fast treatment of different types of cells with different electrical treatments, i.e. different values of electric field parameters.
  • the invention further provides the possibility of application of electric pulses in several different directions thus increasing the membrane area of cell membranes being electroporated.
  • Fig. 1 is a schematic showing a sectional view through a tip electrode chamber on a pipetter 4. Electrodes 2 are mounted into the inner side of a housing 1 and are electrically connected to exterior 3. Fig. 2 is a cross sectional view of the same tip electrode chamber.
  • Fig.3 shows possible directions of the electric field in the lumen of the tip electrode chamber in one case of the position of the electrodes.
  • Fig.4 shows the tip electrode chamber partially filled with a material 6 of the conductance that is comparable to the conductance of the medium of the cell suspension.
  • Tip electrode chamber is designed from housing 1, electrodes 2 and electrical connection 3 to exterior.
  • the housing 1 is made of a non conducting material such as PolyMethylMethAcrylate (PMMA) or other plastic materials.
  • Electrodes 2 and electrical connections 3 are made of electrically conductive material such as metals or their alloys or graphite for example.
  • Tip electrode chamber has his upper part Ia of housing 1 designed so that it fits to pipetter 4.
  • Pipetter 4 usually sucks from 1 ⁇ l to 1000 ⁇ l of fluid.
  • Pipetter 4 is however not an object of this invention.
  • the outer form of lower part Ib of housing 1 is conical, so that it is easier to use with standard laboratory equipment such as centrifuge tubes.
  • Through whole housing 1 in the middle there is an opening 5.
  • cell suspension is sucked into the tip electrode chamber and after electroporation blown out from the tip electrode chamber with pipetter 4. When cell suspension is sucked in it is placed in the opening where electrodes 2 are present. Opening 5 between the electrodes defines the maximum volume of uniformly treated cells.
  • the maximum volume of cell suspension that can be inserted is thus in a range limited by possibilities of construction.
  • the construction allows maximum volumes from 10 ⁇ l to 2500 ⁇ l, predominately from 10 ⁇ l to 1000 ⁇ l.
  • actual volume of uniformly treated cells can be chosen adequately for every experiment simply by adjusting the pipetter.
  • Electrodes 2 are parallelly and symmetrically mounted into the inner surface of the housing 1 and longitudinally to the opening 5.
  • the construction allows length of the electrodes 2 from 5 mm to 60 mm, predominately from 10 mm to 40 mm.
  • Number of electrodes 2 can vary from 2 to 6 electrodes; the said number may be pair or odd.
  • the number of electrodes 2 influence on the number of different directions of electric field that can be used. For example, four electrodes 2 positioned in corners of a quadrant allows for application of pulses in eight different directions (AB and DC, BA and CD, AC, CA, BD, DB, AD and BC, DA and CB). The directions are shown in figure 3a, 3b and 3c. With the number of electrodes the number of different directions of electric field increases.
  • a cross sectional shape of the electrodes 2 There are 3 main options of cross sectional shape of the electrodes 2: rectangular, circular or from one side elliptical, parabolic or hyperbolic and from the other side arbitrary shape.
  • shape of the electrodes is preferably rectangular to get the most homogeneous electric field in the opening 5, but also other aforementioned shapes can be used.
  • shape of the electrodes is usually circular, because they are easier to be produced, but also elliptical, parabolic or hyperbolic one-side shapes can be used, to get more homogeneous electric field in the opening 5, predominately hyperbolic one-side shape.
  • rectangular shape can be used, only that homogeneity of the electric field is very low.
  • Electrodes 2 of rectangular shape can be positioned symmetrically into the centers or preferably for 3 or more electrodes 2 into the corner points of the edge of the equilateral figure of the opening 5.
  • portion of molded surface into the material of the housing 1 depends on the number of electrodes 2.
  • 2 circular electrodes 2 their centers are positioned symmetrically in the center of the edge of the cross sectional shape of the opening 5 and preferably 180° of the electrodes 2 is molded into the material of the housing 1 and preferably 180° of the electrodes 2 is projected out of the material of the housing 1.
  • 3 or more circular electrodes 2 their centers are positioned in the corner points of the cross sectional shape of the opening 5 and their parts, which are projected out of the material of the housing 1, are preferably the same as the inner angles of equilateral figure of the opening 5.
  • Electrodes 2 of circular shape preferably 60° of the electrodes 2 is projected out of the material of the housing 1 and preferably 300° of the electrodes 2 is molded into the material of the housing 1
  • 4 electrodes 2 of circular shape preferably 90° of the electrodes 2 is projected out of the material of the housing 1 and preferably 270° of the electrodes 2 is molded into the material of the housing 1 and so on.
  • elliptical, parabolic or hyperbolic one-side shapes these specific shapes must be oriented into the center of cross sectional shape of the opening 5.
  • the arbitrary shape of elliptical, parabolic or hyperbolic one-side shapes of electrode 2 is preferably completely molded into the material of the housing 1.
  • this arbitrary shape is usually a T profile to get a strong contact between the electrodes 2 and housing 1.
  • Elliptical, parabolic or hyperbolic one-side shapes of the electrodes 2 are preferably all projected out of the material of the housing 1.
  • 2 elliptical, parabolic or hyperbolic one-side shapes of electrodes 2 are used, they are usually positioned symmetrically in the center of the edge of the cross sectional shape of the opening 5.
  • 3 or more elliptical, parabolic or hyperbolic one-side shapes of electrodes 2 are usually positioned in the corner points of the cross sectional shape of the opening 5.
  • the diameter of the electrodes 2 and the distance between them influence on the homogeneity of the electric field in the opening 5. This influence is not so obvious when 2 electrodes 2 of rectangular shape are used.
  • the ratio between the diameter and the distance between the electrodes 2 is optional.
  • the diameter can vary from 0.5 to 7 mm and the distance between them can vary from 0.5 to 6 mm.
  • the choice depends on the maximum volume of uniformly treated cells and on the desired voltage to distance ratio. As aforementioned, the maximum volume of uniformly treated cells is defined as the volume of opening 5 between the electrodes and it is so depended on the length, the diameter and the distance between the electrodes 2.
  • Desired voltage to distance ratio above all depends on cell type and the aim of electroporation and it is usually determined experimentally. Desired voltage to distance ratio vary from 200 V/cm to 4000 V/cm, predominately from 500 V/cm to 1500 V/cm. Generators for electroporation usually generate electrical pulses up to 1000 V. Thus, the distance between the electrodes 2 must be designed so that desired voltage to distance ratio can be achieved. As an example, if used generator for electroporation generates electrical pulses up to 200 V and the desired voltage to distance ratio is 1000 V/cm, the distance between the electrodes 2 must not be greater than 2 mm.
  • the diameter can vary from 0.5 to 5 mm and the distance between them can vary from 0.5 to 6 mm. Ratio between the diameter and the distance between the electrodes 2 of circular shape should be as large as possible to get the most homogeneous electric field in the opening 5.
  • linear conductivity of the shortest line of force between electrically opposite electrodes 2 should be the same as linear conductivity of the line of force between electrically opposite electrodes 2, which goes through the centre of the cross sectional shape of the opening 5, to get the most homogeneous electric field.
  • Linear conductivity is an integral of the current density divided by electric field over the line, which is in our case the line of force.
  • Electrically opposite electrodes are the electrodes 2 which are during electroporation charged with different electrical potential. Electrodes 2 are usually symmetrically charged with different electrical potential. For 3 electrodes 2 of rectangular shape the ratio between the diameter and the distance between the nearest electrodes 2 should be approximately 0.3. The distance between the nearest electrodes 2 can vary from 0.25 to 3 mm.
  • the ratio between the diameter and the distance between the opposite electrodes 2 should be approximately 0.3.
  • the distance between the opposite electrodes 2 can vary from 0.5 to 6 mm.
  • the ratio between the diameter and the distance between the nearest electrodes 2 should be approximately 0.5.
  • the distance between the nearest electrodes 2 can vary from 0.25 to 3 mm.
  • the ratio between the diameter and the distance between the opposite electrodes 2 should be approximately 0.2.
  • the distance between the opposite electrodes 2 can vary from 0.5 to 6 mm.
  • the diameter and the distance between the nearest electrodes 2 should be approximately 1.2.
  • the distance between the nearest electrodes 2 can vary from 0.25 to 2.5 mm.
  • the ratio between the diameter and the distance between the opposite electrodes 2 should be approximately 0.7.
  • the distance between the opposite electrodes 2 can vary from 0.5 to 5 mm.
  • the ratio between the diameter and the distance between the nearest electrodes 2 should be approximately 1.
  • the distance between the nearest electrodes 2 can vary from 0.25 to 2.5 mm.
  • the ratio between the diameter and the distance between the opposite electrodes 2 should be approximately 0.3.
  • the distance between the opposite electrodes 2 can vary from 0.5 to 5 mm.
  • elliptical, parabolic or hyperbolic one-side shapes of electrodes 2 When elliptical, parabolic or hyperbolic one-side shapes of electrodes 2 are used, the ratio between the parameters of one-side shape and the distance between the electrodes 2 is determined numerically. Circular shape of the electrodes 2 is used for the base in the numerical model. Elliptical, parabolic or hyperbolic one-side shape is then appended on the circular shape of numerical model so that the integral of " Manhattan distance" between the two shapes in polar coordinates over the shape, which will be in the direct contact with the cell suspension, is the smallest. Housing 1 and opening 5 with cell suspension is with material conductivity added into the numerical model. Then the electric field between the electrodes 2 of elliptical, parabolic or hyperbolic one-side shape is calculated by finite element method (FEM).
  • FEM finite element method
  • IAE integral absolute error
  • Parameters of the elliptical, parabolic or hyperbolic one-side shape are then iteratively changed so that the estimation of homogeneity of electric field is minimized.
  • Parameters of the elliptical one-side shape are major axis, minor axis and displacement
  • parameters of the parabolic one-side shape are focus and displacement
  • parameters of the hyperbolic one-side shape are semi-major axis, semi-minor axis and displacement.
  • Displacement parameter equals the displacement of the shape from initial position of the shape before minimization of the estimation of homogeneity of electric field. In such a manner obtained shapes are then used for manufacturing of the electrodes 2.
  • design of 3 or more electrodes 2 can also be used in any other electrode chamber for the minimization of inhomogeneous electric field between the electrodes.
  • the ratio between the diameter and the distance between the nearest or opposite electrodes should be the same as previously described for rectangular or circular shape of the electrodes.
  • the same procedure as previously described should be used to determined the most suitable shape of the electrodes regarding the homogeneity of the electrical field between the electrodes.
  • Electrodes are on the top electrically connected to exterior of the tip electrode chamber, so that they can be connected to a generator of electric pulses.
  • Generators usually generate square, sinusoidal and/or exponential electrical pulses up to 1000 V, duration up to 10 s. As aforementioned, generator is not an object of the invention. Electrical connections to exterior are made by curved electrode and/or by additional wire 3.
  • a tip electrode chamber for electroporation according to the invention is characterized in that the opening 5 is in the middle of housing 1 and in the longitudinal direction through the housing 1 and that from two to six electrodes 2 are parallelly and symmetrically mounted into the inner surface of the housing 1 and longitudinally to the opening 5 and partially extending to the said opening 5 and that the said electrodes 2 have electrical connections 3 to exterior.
  • a solid filler 6 can be inserted into the tip electrode chamber or any other electrode chambers as presented on Fig. 4, to minimize inhomogeneous electric field in cell suspension.
  • Solid filler 6 is made from the material of the conductance that is comparable to the conductance of the medium of the cell suspension (0.001 S/cm to 10 S/cm), such as for example certain semiconductor elements (Si, Ge) and their composites or alloys or metal composites or alloys or any other solid materials with the conductance in defined range. Length of the solid filler 6 is preferably the same as the length of the electrodes.
  • Cross sectional shape of the solid filler 6 is preferably the same as the cross sectional shape of the opening, so that solid filler 6 fits into the opening and has a good contact with the electrodes.
  • Solid filler 6 is inserted into the opening and placed between the electrodes.
  • the maximum volume of cell suspension that can be inserted is thus smaller than the opening between the electrodes and it is the same as the volume of the hole 7 described below.
  • Solid filler 6 should be designed so precisely, that it has to be cooled before the insertion. So that it has a good contact with the electrodes and does not fall out after it warms up. To prevent the fall out, solid filler 6 can also be glued on to the housing 1.
  • the diameter of the hole 7 can vary from 1 mm to the diameter of inscribed circle into the cross sectional shape of the solid filler 6 minus 2 mm. Electric field in the hole 7 is in this case almost completely homogeneous since almost all non-homogeneity is distributed inside the solid filler 6 that is on the outer parts of the space between the electrodes.
  • the rectangular shape of electrodes is preferably used, to get a good electrical contact between the electrodes and solid filler.
  • a tip electrode chamber according to the invention is characterized in that the opening 5 is in the middle of housing 1, whereas the solid filler 6 is inserted into the said opening 5 with the hole 7 in the longitudinal direction of the housing 1 and solid filler 6 two to six electrodes 2 are parallelly and symmetrically mounted into the contact of surfaces of the housing 1 and solid filler 6 and that the said electrodes 2 have electrical connections 3 to exterior.
  • the tip electrode chamber is simply attached and detached to the pipetter 4 in the same way as tips used for pipetting. After attaching the tip electrode chamber cell suspension is sucked into the tip electrode chamber and generator is started to generate electrical pulses. During this pulsation electroporation occurs. After the pulsation cell suspension is blown from the tip electrode chamber with the pipetter.
  • the present invention allows electroporation with the aim of electrofusion of cells, gene transfection of cells or electroporation of cells.
  • the operating time for pulsing one volume of cell suspension by this tip electron chamber is minimized because there is much less pipetting needed than with pulsing cells in chambers which have to be filled with a separated pipetter.
  • the filling of the tip electrode chamber is simple with no bubbles in the suspension inserted. Mechanical manipulation is therefore also minimized.
  • the cleaning of the electrode chamber can also be simply done by pulling the cleaning solution up and down several times.
  • the tip can be sterilized by autoclaving or other means depending on the materials used.

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Abstract

La présente invention concerne un appareil d'électroporation conçu avec des pipettes et plusieurs électrodes, avec un champ électrique non homogène minimisé et une charge solide pour les chambres d'électrodes afin de minimiser le champ électrique non homogène. L'invention concerne une chambre d'électrodes de pointe pour électroporation caractérisée en ce que l'ouverture (5) est située au milieu du boîtier (1) et dans la direction longitudinale du boîtier (1), et en ce que de deux à six électrodes (2) sont montées de manière parallèle et symétrique sur la surface interne du boîtier (1), dans la direction longitudinale par rapport à l'ouverture (5), et s'étendent partiellement vers ladite ouverture (5), et en ce que lesdites électrodes (2) présentent des connexions électriques (3) avec l'extérieur. De même, l'invention concerne une chambre d'électrodes de pointe, où la charge solide (6) est insérée dans ladite ouverture (5) munie de l'orifice (7) dans la direction longitudinale du boîtier 1 et où les deux à six électrodes (2) de la charge solide (6) sont montées de manière parallèle et symétrique à l'endroit où les surfaces du boîtier (1) et de la charge solide (6) entrent en contact, et en ce que lesdites électrodes (2) présentent des connexions électriques (3) avec l'extérieur.
PCT/SI2007/000036 2006-10-25 2007-10-03 Chambre d'electrodes de pointe pour electroporation de petit volume WO2008051169A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SI200600250A SI22368A (sl) 2006-10-25 2006-10-25 Konicasta komora z vgrajenimi elektrodami za elektroporacijo manjsega volumna, za elektrofuzijo in gensko transfekcijo
SIP-200600250 2006-10-25

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WO2008051169A1 true WO2008051169A1 (fr) 2008-05-02

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Cited By (7)

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WO2009129327A1 (fr) * 2008-04-15 2009-10-22 Life Technologies Corporation Pointe de pipette pour dispositif d'électroporation
DE102009049783A1 (de) * 2009-10-19 2011-04-21 Eppendorf Ag Elektrisch leitfähige Pipettenspitze
US8383394B2 (en) 2004-06-08 2013-02-26 Applied Biosystems, Llc System and method for electroporating a sample
US20140135198A1 (en) * 2012-11-12 2014-05-15 California Institute Of Technology Devices and methods for nucleic acid extraction capture and concentration using electric fields and centrifugation
US8932850B2 (en) 2004-06-12 2015-01-13 Invitrogen Singapore Pte. Ltd. Electroporation apparatus having an elongated hollow member
US9382510B2 (en) 2011-08-25 2016-07-05 Jian Chen Methods and devices for electroporation
CN110734856A (zh) * 2018-07-20 2020-01-31 苏州壹达生物科技有限公司 一种电击管

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WO2020087074A2 (fr) * 2018-10-26 2020-04-30 Kytopen Corporation Dispositifs, systèmes et kits d'électroporation et leurs procédés d'utilisation

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WO2003057819A1 (fr) * 2002-01-07 2003-07-17 Uab Research Foundation Pointes de pipette de cuvette d'electroporation, reseaux de cuvettes a plusieurs puits, appareil gabarit d'electrode concu pour une automatisation et utilisations de ceux-ci
US6746441B1 (en) * 1998-06-03 2004-06-08 Genetronics, Inc. Flow-through electroporation system for ex vivo gene therapy

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JPS6349070A (ja) * 1986-08-19 1988-03-01 Shimadzu Corp 送液型細胞融合チヤンバ
US6746441B1 (en) * 1998-06-03 2004-06-08 Genetronics, Inc. Flow-through electroporation system for ex vivo gene therapy
WO2003057819A1 (fr) * 2002-01-07 2003-07-17 Uab Research Foundation Pointes de pipette de cuvette d'electroporation, reseaux de cuvettes a plusieurs puits, appareil gabarit d'electrode concu pour une automatisation et utilisations de ceux-ci
DE20302861U1 (de) * 2003-02-21 2003-07-17 Neumann, Eberhard, Prof. Dr., 33739 Bielefeld Biokompatible Elektroden für in vivo Elektroporation zur Wirkstoffzufuhr

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8921092B2 (en) 2004-06-08 2014-12-30 Applied Biosystems Llc System and method for electroporating a sample
US9670449B2 (en) 2004-06-08 2017-06-06 Applied Biosystems Llc System and method for electroporating a sample
US8383394B2 (en) 2004-06-08 2013-02-26 Applied Biosystems, Llc System and method for electroporating a sample
US9260690B2 (en) 2004-06-08 2016-02-16 Applied Biosystems, Llc System and method for electroporating a sample
USRE50241E1 (en) 2004-06-12 2024-12-24 Life Technologies Holdings PTE Ltd. Electroporation apparatus having an elongated hollow member
US8932850B2 (en) 2004-06-12 2015-01-13 Invitrogen Singapore Pte. Ltd. Electroporation apparatus having an elongated hollow member
US11130936B2 (en) 2004-06-12 2021-09-28 Life Technologies Holdings Pte Limited Electroporator apparatus and system having a hollow member
WO2009129327A1 (fr) * 2008-04-15 2009-10-22 Life Technologies Corporation Pointe de pipette pour dispositif d'électroporation
US9523071B2 (en) 2008-04-15 2016-12-20 Life Technologies Corporation Pipette tip for electroporation device
US10450543B2 (en) 2008-04-15 2019-10-22 Life Technologies Corporation Pipette tip for electroporation device
US11634679B2 (en) 2008-04-15 2023-04-25 Life Technologies Corporation Pipette tip for electroporation device
US9346045B2 (en) 2009-10-19 2016-05-24 Eppendorf Ag Electrically conductive pipette tip
DE102009049783A1 (de) * 2009-10-19 2011-04-21 Eppendorf Ag Elektrisch leitfähige Pipettenspitze
US9382510B2 (en) 2011-08-25 2016-07-05 Jian Chen Methods and devices for electroporation
US11959061B2 (en) 2011-08-25 2024-04-16 Jian Chen Methods and devices for electroporation
US20140135198A1 (en) * 2012-11-12 2014-05-15 California Institute Of Technology Devices and methods for nucleic acid extraction capture and concentration using electric fields and centrifugation
CN110734856A (zh) * 2018-07-20 2020-01-31 苏州壹达生物科技有限公司 一种电击管

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