Harberts Jann, Kusch Max, O'Sullivan John, Zierold Robert, Blick Robert H
Center for Hybrid Nanostructures, Universität Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
Department of Physics and Astronomy, University College London, London WC1E 6BT , UK.
Bioengineering (Basel). 2020 May 22;7(2):46. doi: 10.3390/bioengineering7020046.
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.
尽管室温下的膜片钳技术是电生理学界广泛采用的标准程序，但它并不能模拟哺乳动物约37°C的生物系统。为了在电生理研究中更好地模拟自然环境，我们提出了一种为直立显微镜定制的、温度可控的膜片钳平台，该平台可以轻松独立地适配任何直立膜片钳装置，无论是市售的还是自制的。我们的装置能够对平台进行冷却和加热，温度变化仅小于0.5°C。我们通过在36°C下对Jurkat T淋巴细胞和人诱导多能干细胞衍生神经元进行膜片钳测量来展示我们的装置。将被动膜参数和特征电生理特性，如电压门控离子通道的门控特性和动作电位的发放，与室温下的测量结果进行比较。我们观察到，许多未被明确认为与温度相关的过程会随温度变化。因此，我们认为如果需要改善生理条件，膜片钳测量中需要进行温度控制。此外，我们建议研究人员仅直接比较在相似温度下测量的电生理结果，因为温度变化可能会导致细胞特性的微小差异。
