Experimental determination of Cm measurement related hardware parameters of the patch-clamp amplifier

Accurate Cm measurements rely on accurate determination of specific parameters of a patch-clamp amplifier (PCA). Hardware-related parameters, such as the resistance Rf and the stray capacitance Cf of the feedback resistor, the input capacitance Ci, the injection capacitance Cj, and the extra capacitances introduced by the BNC connector, are of significance in the sense of obtaining absolute estimates of cell parameters. In the present paper, a frequency-domain method, or the f-method for simplicity, is put forward to experimentally determine the actual values of basic circuit elements for our self-developed PCA. The f-method makes use of sine waves and amplitude/phase measurements instead of the square-wave responses to determine the above parameters of a PCA, and thereby calibrates the PAC for capacitance measurements. Experimental results prove that the f-method is excellent in determining hardware-related parameters, with 3-5% error of the impedance of the “10 MΩ setting”, and about 2% error of the impedance of the “model cell” of the model circuit for our PCA. The f-method enables us not only to picture components of fast capacitances, but also to guarantee complete fast capacitance compensation; it may be applicable for other PCAs.