PubMed · 4413209
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H E Henkes, G H Van Lith. 1974. Electroperimetry.. https://doi.org/10.1159/000307119
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A novel micropipette holder has been designed to overcome the problem of drift encountered when experimenting on small preparations using patch-clamp techniques. In order to increase the stiffness and thermal stability of the system, the holder is designed such that the micropipette is clamped to a rigid quartz rod, and isolated from stresses arising in the connectors. The pressure line, made from stiff tubing, and electrical leads are firmly attached to the table and micromanipulator. This design was able to hold the micropipette tip stable to with 100 nm for more than 1 h, even with changes of 26.7 kPa (200 mmHg) in the pressure line.
A new method for fast perfusion of single adherent cells during whole-cell and single-channel patch-clamp measurements is described. The main advantages over previous methods are: (1) the solution surrounding a single cell in a Petri dish (or cover slip) can be exchanged without contamination of the entire cell population, and (2) only small quantities of test solution (50 microliters) are required. The method consists of inserting a patch pipette into a micro-perfusion chamber (MPC) where whole-cell and single-channel currents can be studied. A small volume (2-3 microliters) surrounding adherent single cells in a Petri dish is separated by enclosing it in a ring of Silicone rubber which is then pressed to the bottom of the dish. A drug-containing test solution (50 microliters) is supplied to a funnel at the chamber inflow. The MPC volume isolated from the main solution in the Petri dish is rapidly changed when suction is applied to the chamber outflow. The speed of solution exchange in the MPC (16 +/- 5 ms, n = 12) was estimated by observing changes in the tip potential of 1-M omega patch pipettes during rapid chamber perfusion with solutions of different ionic composition. The seal of the MPC with the surface of a Petri dish was approximated by measuring the electrical resistance between the MPC interior and a reference electrode placed in the Petri dish outside the MPC (range 300-500 M omega). Additionally, a radioactive calcium channel ligand [3H]isradipine was added to the MPC and the appearance of radioactivity in the Petri dish (outside the MPC) subsequently measured.(ABSTRACT TRUNCATED AT 250 WORDS)
Differential input amplifiers are commonly used in the measurement of biopotential signals. This is because of their ability to reject common-mode signals, for example power-line interference. Rejection is further enhanced by referencing the subject to the instrumentation system. One such referencing technique uses a third subject electrode connected either to circuit common or to ground. However, these techniques are rarely explained or quantified in the literature. Four recording configurations are examined, isolated and non-isolated amplifiers with two and with three electrodes, and existing models are extended. From a brief assessment of power-line interference sources, it is concluded that only displacement currents induced into the leads and the body need be considered. The effect of these on the four configurations is analysed using SPICE. Results show that interference from displacement currents induced into the leads and into the body is dependent upon the recording configuration used and is significantly lower in three-electrode than in two-electrode amplifiers. Isolation also reduces levels of interference. It is concluded that the choice of referencing may affect interference levels on the biopotential signal significantly. Experimental and further analytical work is planned to confirm this.