Highly controllable electroporation and applications thereof

Abstract

The controllable electroporation system and method described herein allows control over the size, the number, the location, and the distribution of aqueous pores, thus increasing flexibility of use. The herein described system and method for controllable electroporation generally employs at least two actuating sub-systems and sub-processes. One sub-system and sub-process employs a relatively broad effect in order to weaken the membrane, a broad effect sub-system. Another sub-system and sub-process employs a relatively narrow effect in order to localize the position of the pore in the membrane, a narrow effect sub-system.

1 . A system for controllable electroporation of a membrane comprising: 
 a broad energy sub-system operably coupled to the membrane and    a narrow energy sub-system operably coupled to the membrane,    wherein a pore is opened or created at a position corresponding to the position of the narrow energy.    
     
     
         2 . The system as in  claim 1 , wherein the broad energy sub-system is selected from the group of weakening systems consisting of electric fields, microwave energy, other electromagnetic radiation, low energy laser beams, or any combination comprising at least one of the foregoing weakening systems.  
     
     
         3 . The system as in  claim 1 , wherein the energy magnitude of the broad energy sub-system is lower than the energy magnitude of electroporation systems without the narrow effect sub-system whereby random pore opening occur.  
     
     
         4 . The system as in  claim 1 , wherein the area of the broad energy sub-system encompasses an area larger than the desired pore size.  
     
     
         5 . The system as in  claim 1 , wherein the area of the broad energy sub-system encompasses the membrane of a cell.  
     
     
         6 . The system as in  claim 1 , wherein the area of the broad energy sub-system encompasses membranes of an array of cells.  
     
     
         7 . The system as in  claim 1 , wherein the area of the broad energy sub-system encompasses a region of a membrane.  
     
     
         8 . The system as in  claim 1 , wherein the narrow energy sub-system is selected from the group of position localization systems consisting of laser beams, electrode tips, or any combination comprising at least one of the foregoing position localization systems.  
     
     
         9 . The system as in  claim 1 , wherein the area of the narrow energy sub-system corresponds to the dimensions of the pore opening.  
     
     
         10 . The system as in  claim 1 , wherein the pore has sub-micron dimensions.  
     
     
         11 . The system as in  claim 1 , wherein the pore has dimensions of about 100 nanometers or less.  
     
     
         12 . A controllable electroporation system comprising: 
 a broad energy sub-system operably coupled for providing broad energy to the membrane and    a narrow energy sub-system operably coupled to the membrane,    wherein a pore is opened when both the broad energy sub-system and the narrow energy sub-system are activated.    
     
     
         13 - 23 . (canceled)  
     
     
         24 . A cell pore opening system comprising: 
 a microrobotic device for holding a cell and    a system as in  claim 1  for controllably opening a pore in the cell.    
     
     
         25 . The cell pore opening system as in  claim 24 , wherein the broad energy sub-system comprises an electrode plate and a switchable voltage source.  
     
     
         26 . The cell pore opening system as in  claim 24 , wherein the narrow energy sub-system comprises a laser.  
     
     
         27 . A cell pore macromolecule system comprising: 
 a microrobotic device for holding a cell;    a system as in  claim 1  for controllably opening a pore in the cell; and    a macromolecule injection device for injecting a macromolecule into the cell via the pore.    
     
     
         28 - 29 . (canceled)  
     
     
         30 . A system for filtering molecules or macromolecules comprising: 
 a plurality of membrane layers, each membrane layer including a system as in  claim 1  for controllably opening a pore in the cell, each membrane layer opened to a different size to create a pore size gradient.    
     
     
         31 . A system for filtering molecules or macromolecules comprising: 
 a plurality of membrane layers,    a broad energy sub-system associated with each membrane layer,    wherein each layer includes a position having a defect whereby said position is closed without activation of the broad energy sub-system and said position is opened upon activation of the broad energy sub-system.    
     
     
         32 . The system as in  claim 31 , wherein said defect at each layer controls the size of the pore.  
     
     
         33 . The system as in  claim 31 , wherein the magnitude of the energy of the broad energy sub-system controls the size of the pore.  
     
     
         34 . A system for filtering molecules or macromolecules comprising: 
 a membrane layer including a system as in  claim 1  for controllably opening a pore in the cell,    wherein the narrow energy sub-system comprises an array of narrow energy sub-sub-systems.    
     
     
         35 . A system for filtering molecules or macromolecules comprising: 
 a membrane layer including a system as in  claim 1  for controllably opening a pore in the cell,    wherein the narrow energy sub-system comprises an array of lasers.    
     
     
         36 . A system for filtering molecules or macromolecules comprising: 
 a membrane layer including a system as in  claim 1  for controllably opening a pore in the cell,    wherein the narrow energy sub-system comprises a beam steering device associated with a source of electromagnetic energy.    
     
     
         37 - 47 . (canceled)