A COMBINED ELECTROPORATION AND MICROINJECTION METHOD FOR THE PENETRATION OF LIPID BILAYER MEMBRANES

摘要

Opisan je postupak za penetraciju dvoslojnih lipidnih membrana kako bi se ubacio vrh upljeg objekta u obliku igle, kao to je mikropipeta u komoru koji je obrazovan od dvoslojne lipidne membrane, pri emu je data komora tako smetena, da se nalazi izmedju pomenutog objekta u obliku igle, sa vrhom koji je u kontaktu sa komorom tako da primenjuje mehaniku silu na lipidnu membranu komore, te je na taj nain komora pod naponom, i druge elektrode, pri emu je uspostavljen prolazni elektrini puls 1 do 103V/cm izmedju dve eklektrode, to je rezultovalo u formiranju fokusiranog elektrinog polja du komore, koje dalje indukuje dielektrino raspadanje lipidnog dvosloja, to omoguava penetraciju objekta u obliku igle u kontejner. Takodje je opisan postupak elektroinjektiranja koji je zasnovan na prethodno opisanom postupku, pri emu je supstanca ubaena kroz objekat u obliku igle i kontejner, posle penetracije objekta u obliku igle.[Disclosed is a method for penetration of lipid bilayer membranes in order to insert at the tip of a hollow needle-shaped object, such as a micropipet-, into a container formed of a lipid bilayer membrane, wherein said container is placed between said needle-shaped object, with the tip of said needle-shaped object placed in contact with said conainer in such a way that it applies a mechanical force to the lipid membrane of said container, thus mechanically straining it, and a second electrode, whereupon a transient electric pulse of 1-to-103 V/cm is applied between the electrodes, resulting in a focused electrical field over said container which induces a dielectric breakdown of the lipid bilayer causing the needle-shaped object to penetrate the container. Disclosed is also an electroinjection method based on the above method, wherein substances are introduced through the needle-shaped object and into the container after penetration of the needle-shaped object.;Disclosed is a method for penetration of lipid bilayer membranes in order to insert at the tip of a hollow needle-shaped object, such as a micropipet-, into a container formed of a lipid bilayer membrane, wherein said container is placed between said needle-shaped object, with the tip of said needle-shaped object placed in contact with said conainer in such a way that it applies a mechanical force to the lipid membrane of said container, thus mechanically straining it, and a second electrode, whereupon a transient electric pulse of 1-to-103 V/cm is applied between the electrodes, resulting in a focused electrical field over said container which induces a dielectric breakdown of the lipid bilayer causing the needle-shaped object to penetrate the container. Disclosed is also an electroinjection method based on the above method, wherein substances are introduced through the needle-shaped object and into the container after penetration of the needle-shaped object.