Adjusting the distance of electrochemical microsensors from secreting cell monolayers on the micrometer scale using...

There are emerging applications of electrochemical microsensors where the distance of the sensor from an insulating plane needs to be adjusted and/or accurately known. The plane may be merely an obstruction or the source of a species whose release rate needs to be measured. An example is in cell secretion studies where a monolayer of cells is stimulated to secrete ions and/or other biochemical species which then diffuse away from the cells while being measured by a microsensor. Sensor response will thus depend on both the rate of release and the distance of the sensor from the cells. To obtain accurate release rates, the precision of the scheme to control electrode distance from the monolayer needs to be on the micrometer scale for species with ionic diffusivities. Optical (stereomicroscope and microruler) and mechanical (precalibrated micrometer screw) methods to precisely position the electrode are difficult to use under realistic circumstances (due to opaqueness of the chamber, and/or the medium, or irreproducible chamber depth). In this work we propose to correlate electrochemical cell impedance with sensor distance. This scheme has been used to adjust the distance of a chloride (tip diameter approximately 250 microns) and a potassium (tip diameter approximately 1000 microns) ion-selective microelectrode in the 0-250- and 0-2500-micron range, respectively, from a planar obstruction as well as from a monolayer of cells with a best precision of +/- 5 microns (n = 6) for the chloride and about +/- 20 microns for the potassium sensor. Larger electrodes have a broader range of distances over which they are sensitive, albeit with a poorer spatial resolution. This was verified by using Ag disk electrodes of 250 and 500 microns in diameters in AgNO3 solution.