Brüggemann A, George M, Klau M, Beckler M, Steindl J, Behrends J C, Fertig N
Nanion Technologies GmbH, München, Germany.
Assay Drug Dev Technol. 2003 Oct;1(5):665-73. doi: 10.1089/154065803770381020.
In evaluating ion channel function, electrophysiology, e.g., patch clamping, provides the highest information content. For the analysis of ion channel-modulating compounds, one variant of the patch-clamp technique, the whole-cell configuration, is particularly useful. We present here patch-clamp recordings in the whole-cell configuration and single channel recordings performed with planar patch-clamp chips, which are microstructured from borosilicate glass substrate. The chips are used in the Port-a-Patch, an ion channel research/screening instrument that enables automated patch-clamp experiments on a single cell. A software runs the experiment by executing user-determined protocols for cell positioning, as well as for electrical stimulation and current readout. In various electrophysiological experiments, the high quality of recordings and the versatility of the perfusion of the recorded cells are demonstrated. Quantitative pharmacological experiments are performed on sodium channels expressed in HEK cells using solution volumes in the low microliter range. The exceptionally low volume consumption in the experiments make the system attractive for work on rare or expensive compounds. Due to the low volumes necessary, a rapid solution exchange is facilitated, which is shown on RBL cells. The patch-clamp chip enables a rapid and precise perfusion, allowing sophisticated investigations on ion channel function with the Port-a-Patch.
在评估离子通道功能时,电生理学方法,例如膜片钳技术,能提供最丰富的信息。对于离子通道调节化合物的分析,膜片钳技术的一种变体——全细胞模式尤其有用。我们在此展示全细胞模式下的膜片钳记录以及使用由硼硅酸盐玻璃基板微结构化制成的平面膜片钳芯片进行的单通道记录。这些芯片用于Port-a-Patch,这是一种离子通道研究/筛选仪器,可在单个细胞上进行自动化膜片钳实验。软件通过执行用户确定的细胞定位协议以及电刺激和电流读出协议来运行实验。在各种电生理实验中,展示了记录的高质量以及所记录细胞灌注的多功能性。使用低微升范围内的溶液体积,对HEK细胞中表达的钠通道进行了定量药理学实验。实验中极低的溶液消耗量使该系统对于处理稀有或昂贵化合物的工作具有吸引力。由于所需体积小,促进了溶液的快速交换,这在RBL细胞上得到了证明。膜片钳芯片能够实现快速精确的灌注,从而可以使用Port-a-Patch对离子通道功能进行深入研究。