Ion channel assay methods
View Patent ↗A method of characterizing the biological activity of a candidate compound may include exposing cells to the candidate compound, and then exposing the cells to a repetitive application of electric fields so as to set the transmembrane potential to a level corresponding to a pre-selected voltage dependent state of a target ion channel.
1. A method of characterizing the biological activity of a candidate compound comprising:
placing one or more cells into an area of observation in a sample well;
exposing said one or more cells to said compound;
repetitively exposing said one or more cells to a series of biphasic electric fields at a rate of approximately 20 to 100 pulses per second;
producing a limited spatial variation in intensity in said electric field in said area of observation of within ±10% from a mean intensity in that area;
producing a controlled change in transmembrane potential predominantly in a single direction away from a starting transmembrane potential during said series of biphasic electric fields due to a continuing and additive accumulation of charge in said one or more cells over the course of said series of electric fields; and
detecting said controlled change in transmembrane potential of said one or more cells by detecting fluorescence emission of a FRET based voltage sensor from an area of observation containing said one or more cells.
2. The method of claim 1 , wherein said one or more electrical fields cause an ion channel of interest to open.
3. The method of claim 1 , wherein said one or more electrical fields cause an ion channel of interest to be released from inactivation.
4. The method of claim 1 , wherein said one or more cells comprise a voltage regulated ion channel.
5. The method of claim 4 , wherein said voltage regulated ion channel is selected from the group consisting of a potassium channel, a calcium channel, a chloride channel and a sodium channel.
6. The method of claim 1 , wherein spatial variation in said one or more electrical fields is within ±1% from the mean electrical field at any one time.
7. The method of claim 1 , wherein said one or more electrical fields are selected from a square wave-form, a sinusoidal wave-form or a saw tooth wave-form.