Department of Mathematics and Natural Sciences, Carl-von-Ossietzky University Oldenburg, Oldenburg, Germany.
J Neurosci Methods. 2013 Sep 30;219(1):83-91. doi: 10.1016/j.jneumeth.2013.06.007. Epub 2013 Jul 5.
Since temperature severely affects all physiological processes, exact temperature control during electrophysiological measurements is indispensable. However, none of the tempering system approaches previously described is fully satisfactory for extracellular recordings with sharp multi-electrode arrays (MEAs).
We developed a set-up offering a homogeneously tempered and at the same time light-transparent stage for an ex vivo preparation. The Peltier element based tempering unit of our system is physically separated from the preparation stage avoiding electrical disturbances of extracellular recordings. We implemented a digital feedback controller on a microcontroller to minimise the deviation between actual and set point temperature.
Our tempering system allows operation from 10°C to 45°C with a control error in steady state between 0.052°C (RMSE) and 0.115°C (RMSE). To document the versatility of our system, we performed extracellular MEA recordings from retinal ganglion cells of isolated retina under different temperature conditions. We found strong influences on light response properties, even for small temperature changes.
Currently used heating systems that allow top and bottom side optical access to a preparation typically exhibit low temperature accuracy, precision or homogeneity.
Our system is adequate not only for experiments on a variety of species under physiological temperature conditions but also for studies on temperature effects on physiology in general. Though the setup was developed for the context of MEA recordings from retina it may be useful in other cases where optical access to the preparation from both, top and bottom side is required.
由于温度严重影响所有生理过程,因此在电生理测量过程中进行精确的温度控制是必不可少的。然而,以前描述的调温系统都不能完全满足具有尖锐多电极阵列 (MEA) 的细胞外记录的要求。
我们开发了一种设置,为离体准备提供均匀调温和同时透光的阶段。我们系统的基于 Peltier 元件的调温单元与准备阶段物理分离,避免了细胞外记录的电干扰。我们在微控制器上实现了数字反馈控制器,以最小化实际温度和设定点温度之间的偏差。
我们的调温系统允许在 10°C 到 45°C 之间操作,在稳态下控制误差在 0.052°C(均方根误差)和 0.115°C(均方根误差)之间。为了证明我们系统的多功能性,我们在不同温度条件下从离体视网膜的神经节细胞进行了细胞外 MEA 记录。我们发现即使温度变化很小,也会对光反应特性产生强烈影响。
目前允许对准备进行顶部和底部光学访问的加热系统通常表现出低温度精度、精度或均匀性。
我们的系统不仅适用于在生理温度条件下对各种物种进行的实验,也适用于一般研究温度对生理学的影响。尽管该设置是为从视网膜进行 MEA 记录的背景而开发的,但它可能在其他需要从顶部和底部对准备进行光学访问的情况下有用。