Strauss U, Herbrik M, Mix E, Schubert R, Rolfs A
Department of Neurology, University of Rostock, Germany.
Pflugers Arch. 2001 Jul;442(4):634-8. doi: 10.1007/s004240100578.
Perforated whole-cell patch-clamp recordings obtained with nystatin are frequently used to preserve intracellular integrity. However, the perforated-patch configuration may sometimes undergo a spontaneous change into the conventional whole-cell configuration, especially when lymphocytes are investigated. The electrophysiological criteria-- previously described--for establishing the existence of the perforated whole-cell configuration have been shown to be insufficient. Thus, the dye eosin, applied to the pipette solution, was tested as a tool for discriminating between the perforated and the conventional whole-cell configurations on rat T-lymphocytes. The dye never entered the cell from the pipette during the entire measurement in the perforated whole-cell configuration. In contrast, all cells in the conventional whole-cell configuration became red immediately after membrane rupture. Eosin barely changed the currents studied. The results suggest that eosin is a dye of choice for verifying a true perforated-patch configuration.
制霉菌素穿孔全细胞膜片钳记录常用于维持细胞内完整性。然而，穿孔膜片钳配置有时会自发转变为传统全细胞膜片钳配置，特别是在研究淋巴细胞时。先前描述的用于确定穿孔全细胞膜片钳配置存在的电生理标准已被证明是不够的。因此，将添加到移液管溶液中的染料伊红作为一种区分大鼠T淋巴细胞穿孔和传统全细胞膜片钳配置的工具进行了测试。在穿孔全细胞膜片钳配置的整个测量过程中，染料从未从移液管进入细胞。相比之下，传统全细胞膜片钳配置中的所有细胞在膜破裂后立即变红。伊红对所研究的电流几乎没有影响。结果表明，伊红是验证真正穿孔膜片钳配置的首选染料。
