System and method for obtaining and maintaining high-resistance seals in patch clamp recordings

Abstract

The invention provides a system, system components, and a method for rapidly obtaining and stably maintaining a cell in optimal contact with the cell-contacting surface of a sensor in a cell-based biosensor. In one aspect, the system maximizes the seal between a whole cell and the cell-contact surface of a patch clamp micropipette, maximizing the efficiency of a whole cell patch clamp recording.

1 - 73 . (canceled) 
   
   
       74 . A method for producing an electrically resistant seal between a cell and a cell-contacting surface comprising exposing the cell to at least one fluid stream. 
   
   
       75 . The method of  claim 74 , wherein electrical resistance is at least about 100 Mohm. 
   
   
       76 . The method of  claim 74 , further comprising measuring electrical properties of the cell. 
   
   
       77 . The method of  claim 74 , wherein the stream is a u-shaped fluid stream. 
   
   
       78 . The method of  claim 74 , further comprising measuring the properties before and after exposing the cell to an agent. 
   
   
       79 . The method of  claim 78 , wherein the agent is selected from the group consisting of: protein, DNA, RNA, PNA, a receptor agonist, a receptor antagonist, a neurotransmitter, a neurotransmitter analog, an enzyme inhibitor, an ion channel modulator, a G-protein coupled receptor modulator, a transport inhibitor, a hormone, a peptide, a toxin, an antibody, a pharmaceutical agent, a chemical, a purinergic compound, a cholinergic compound, a serotonergic compound, a dopaminergic compound, an anesthetic, a benzodiazepine, a barbiturates, a steroid, an alcohol, a metal cation, a cannabinoid, a cholecystokinin, a cytokine, an excitatory amino acid, a GABAergic agent, a ganglioside, a histaminergic compound, a melatonin, a neuropeptide, a neurotoxin, an endothelins, an NO compound, an opioid, a sigma receptor ligand, a somatostatin, a tachykinin, an angiotensin, a bombesin, a bradykinin, a prostaglandin and combinations thereof. 
   
   
       80 . The method of  claim 74 , wherein the method further comprises providing a substantially planar substrate in communication with at least one conducting element, wherein the substantially planar structure comprises at least one nonplanar element for establishing electrical communication with a cell and which comprises the cell-contacting surface. 
   
   
       81 . The method of  claim 74 , wherein the method further comprises applying a voltage pulse at the cell-contacting surface to enhance electrical resistance of the seal. 
   
   
       82 . The method of  claim 74 , wherein the method further comprises applying pressure at the cell-contacting surface to enhance electrical resistance of the seal. 
   
   
       83 . The method of  claim 76 , wherein the method comprises providing a plurality of fluid streams. 
   
   
       84 . The method of  claim 83 , wherein two or more of the fluid streams contact the cell and one or both streams delivers an agent to the cell. 
   
   
       85 . The method of  claim 83 , wherein two or more of the fluid streams contact the cell and at least two streams deliver different agents or different concentrations of the same agent. 
   
   
       86 . The method of  claim 84 , wherein at least one stream delivers a buffer to the cell. 
   
   
       87 . The method of  claim 74 , wherein the method further comprises providing a substantially planar substrate in communication with at least one conducting element, wherein the substantially planar structure comprises at least one nonplanar element for establishing and/or maintaining electrical communication with a cell, wherein the at least one nonplanar element comprises a portion of a wall. 
   
   
       88 . The method of  claim 74 , wherein the method further comprises providing a substantially planar substrate in communication with at least one conducting element, wherein the substantially planar structure comprises at least one nonplanar element for establishing and/or maintaining electrical communication with a cell, wherein the positioner comprises a protruded surface of a cell chamber. 
   
   
       89 . The method of  claim 87 , wherein the nonplanar element comprises one or more of a protruded surface defining an opening or a portion of a wall. 
   
   
       90 . The method of  claim 87 , wherein the protruded surface defining an opening comprises one or more of a microchannel, a column, a pyramidal element, rod or reeve. 
   
   
       91 . The method of  claim 87 , wherein the electrical resistance is at least about 100 Mohm. 
   
   
       92 . The method of  claim 87 , wherein the method further comprises measuring electrical properties of the cell.