Improved superfusion technique for rapid cooling or heating of cultured cells under patch-clamp conditions

I Dittert, J Benedikt, L Vyklický, K Zimmermann… - Journal of neuroscience …, 2006 - Elsevier
I Dittert, J Benedikt, L Vyklický, K Zimmermann, PW Reeh, V Vlachová
Journal of neuroscience methods, 2006Elsevier
We have developed an improved technique for fast cooling and heating of solutions
superfusing isolated cells under patch-clamp or calcium imaging conditions. The system
meets the requirements for studying temperature dependency of all kinds of ion channels, in
particular temperature-gated ion channels. It allows the application of temperature changes
within a range of 5–60° C at maximum rates of− 40° C/s to 60° C/s. Barrels filled with
different solutions are connected to a manifold consisting of seven silica capillaries (320μm …
We have developed an improved technique for fast cooling and heating of solutions superfusing isolated cells under patch-clamp or calcium imaging conditions. The system meets the requirements for studying temperature dependency of all kinds of ion channels, in particular temperature-gated ion channels. It allows the application of temperature changes within a range of 5–60°C at maximum rates of −40°C/s to 60°C/s. Barrels filled with different solutions are connected to a manifold consisting of seven silica capillaries (320μm inner diameter, i.d.). A common outlet consists of a glass capillary through which the solutions are applied onto the cell surface. The upper part of this capillary is embedded in a temperature exchanger driven by a miniature Peltier device which preconditions the temperature of the passing solution. The lower part of the capillary carries an insulated copper wire, densely coiled over a length of 7mm, and connected to a dc current source for resistive heating. The Peltier device and the heating element are electrically connected to the headstage probe which is fixed on to a micromanipulator for positioning of the manifold. The temperature of the flowing solution is measured by a miniature thermocouple inserted into the common outlet capillary near to its orifice which is placed at a distance of less than 100μm from the surface of the examined cell. The temperature is either manually controlled by voltage commands or adjusted via the digital-to-analog converter of a conventional data acquisition interface. Examples are given of using the device in patch-clamp studies on heterologously expressed TRPV1, TRPM8, and on cultured rat sensory neurons.
Elsevier