Abstract
Though patch clamping at room temperature is a widely disseminated standard procedure in the electrophysiological community, it does not represent the biological system in mammals at around 37 °C. In order to better mimic the natural environment in electrophysiological studies, we present a custom-built, temperature-controlled patch clamp platform for upright microscopes, which can easily be adapted to any upright patch clamp setup independently, whether commercially available or home built. Our setup can both cool and heat the platform having only small temperature variations of less than 0.5 °C. We demonstrate our setup with patch clamp measurements at 36 °C on Jurkat T lymphocytes and human induced pluripotent stem cell-derived neurons. Passive membrane parameters and characteristic electrophysiological properties, such as the gating properties of voltage-gated ion channels and the firing of action potentials, are compared to measurements at room temperature. We observe that many processes that are not explicitly considered as temperature dependent show changes with temperature. Thus, we believe in the need of a temperature control in patch clamp measurements if improved physiological conditions are required. Furthermore, we advise researchers to only compare electrophysiological results directly that have been measured at similar temperatures since small variations in cellular properties might be caused by temperature alterations.
Highlights
In 1780, Luigi Galvani set the cornerstone for modern electrophysiology by accidentally discovering electrical processes within living creatures as electrical currents evoked the movement of frog legs [1]
We know that our entire nervous system is based on the transmission of electrical as well as chemical signals on a single-cell scale, and patch clamping helped fundamentally in understanding the role of many different ion channels in the context of physiological and pathophysiological functions [5,6,7]
Na+, K+, and Ca2+ voltage-gated ion channels play an important role in the nervous system, and the accurate apprehension of the electrical signaling could even render a direct connection between the nervous system and a computer [8,9]
Summary
In 1780, Luigi Galvani set the cornerstone for modern electrophysiology by accidentally discovering electrical processes within living creatures as electrical currents evoked the movement of frog legs [1]. In vivo (automated) patch clamping offers the opportunity to study electrophysiology in living animals—a setting as close as it can get to nature—but such applications are even more challenging due to more complex setups and low giga-seal probabilities [14,15]. A patch clamp setup allowing for electrophysiological measurements at controlled temperatures would be of great interest to researchers in order to shed light onto temperature depending cell processes and functions [25]. We present a low-cost, low-tech approach—feasible for everybody and adaptable to every setup independently, whether commercially bought or in-house built—for controlling the temperature of biological samples during patch clamp measurements. We found small influences on non- temperature depending processes confirming the potential need of a temperature control in patch clamp measurements if best biological conditions are required. Researchers should only compare their data to the results measured at same temperatures if differences are highlighted compared to literature
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