Electroporation apparatus having an elongated hollow member

Abstract

An electroporation apparatus comprising an elongated hollow member in order to provide a uniform electric field during electroporation, wherein specifically, electroporation is carried out by applying electric pulses through a couple of electrodes from both end parts of the elongated hollow member, after the hollow member is charged with fluid specimen including cells and material which would be injected into the cells.

What is claimed is: 
     
       1. An electroporation apparatus for applying an electric pulse or electric pulses to a specimen including cells to thereby electroporate cell membranes and infuse foreign materials into the cells, comprising:
 a longan elongated hollow specimen-stuffing member composed of non-conductive material; 
 a pressure maintaining means connected to a distal first end of the elongated hollow specimen-stuffing member for fluid communication; 
 a reservoir connected to the other distal a second end of the elongated hollow specimen-stuffing member for fluid communication and disposed with an electrode for electrically contacting the a specimen or an electrolytic solution disposed in the elongated hollow specimen-stuffing member; and 
 a reservoir holder including a fixing unit for fixing the pressure maintaining means, an a first electrode terminal for electrically connecting the fixing unit to the pressure maintaining means and another a second electrode terminal for electrically connecting the electrode disposed at the reservoir; 
 wherein the pressure maintaining means is a pipette in which a conductive contact is disposed at part of the a pipette body thereof of the pipette and a movable electrode is inserted into the conductive contact for communication with a piston, the movable electrode also being inserted into the specimen stuffing elongated hollow specimen-stuffing member for fluid communication and for electrically contacting the specimen or an the electrolytic solution disposed in the specimen stuffing elongated hollow specimen-stuffing member, the pippette pipette being in electrical connection with the movable electrode through the conductive contact, and 
 wherein the elongated hollow specimen-stuffing member is directly attached and detached to a tip mounting shaft of the pipette. 
 
     
     
       2. The electroporation apparatus according to  claim 1 , wherein the elongated hollow specimen-stuffing member is a capillary or a tubing. 
     
     
       3. The electroporation apparatus according to  claim 1 , wherein the elongated hollow specimen-stuffing member has a ratio (R, cm −1 ) of a longitudinal length (L, cm) to horizontal cross-sectional area (A, cm 2 ) in the range of 50 to 10,000. 
     
     
       4. The electroporation apparatus according to  claim 1 , wherein the movable electrode is a plastic of which a surface is coated with conductive material. 
     
     
       5. An electroporation system for introducing foreign materials into cells by eletroporating electroporating cell membranes by way of applying an electric pulse or electric pulses to a specimen including the cells, comprising:
 the electroporation apparatus according to  claim 1 ; and 
 a pulse generator for generating an electric pulse, 
 wherein the electrode disposed in the reservoir is disposed with an electrode contactingis in electrical contact with the specimen or anthe electrolytic solution, the elongated hollow specimen-stuffing member is filled with the specimen by the pressure maintaining means, the specimen or the electrolytic solution filled in the reservoir is connected to a distal the second end of the elongated hollow specimen-stuffing member for fluid communication, and an electric pulse or electric pulses are applied to anthe second electrode contacting the specimen or the electrolytic solution filled in the reservoir and anotherthe first electrode being inserted into an electrode insertion unit of theof a connector to thereby electroporate the cells in the specimen filled in the elongated hollow specimen-stuffing member. 
 
     
     
       6. An electroporation system for introducing foreign materials into cells by eletroporating electroporating cell membranes by way of applying an electric pulse or electric pulses to a specimen including the cells, comprising:
 the electroporation apparatus according to  claim 1 ; and 
 a pulse generator for generating an electric pulse, 
 wherein the pressure maintaining means is a pipette disposed at part of the body thereof with a conductive contact, and a the movable electrode is disposed inside the elongated hollow specimen-stuffing member is inserted for communication with a the piston, and 
 wherein the hollow specimen-stuffing member is directly detached and attached to a tip mounted shaft of the pipette, the movable electrode is raised or lowered to a distal the first end or the second end of the elongated hollow specimen-stuffing member by a depression button of the pipette to fill the specimen in the elongated hollow specimen-stuffing member or retrieve it, the pipette is inserted and fixed to a reservoir holder inner pipe wall including the fixing unit, a the conductive contact of the pipette body is electrically connected to the first electrode terminal via the fixing unit of the reservoir holder inner pipe wall, the elongated hollow specimen-stuffing member is so positioned as to fluidly communicate with the specimen or the electrolytic solution stored in the reservoir, and an electric pulse or electric pulses are applied to the electrode contacting the specimen or the electrolytic solution stored in the reservoir to thereby electroporate the cells in the specimen filled in the elongated hollow specimen-stuffing member. 
 
     
     
       7. The electroporation apparatus according to  claim 1 , wherein the reservoir is composed of non-conductive material. 
     
     
       8. The electroporation apparatus according to  claim 7 , wherein the reservoir is transparent. 
     
     
       9. The electroporation apparatus according to  claim 8 , wherein the reservoir is composed of polystyrene. 
     
     
       10. The electroporation apparatus according to  claim 1 , wherein the elongated hollow specimen-stuffing member is transparent. 
     
     
       11. The electroporation apparatus according to  claim 10 , wherein the elongated hollow specimen-stuffing member is composed of polypropylene. 
     
     
       12. The electroporation apparatus according to  claim 1 , the electrode comprises platinum, gold, silver, or copper. 
     
     
       13. The electroporation apparatus according to  claim 1 , wherein the reservoir holder comprises an upper lid and a body. 
     
     
       14. The electroporation apparatus according to  claim 13 , wherein the upper lid and the body are separable. 
     
     
       15. The electroporation apparatus according to  claim 13 , wherein the upper lid and the body are integrally formed.