A new technique for multiple re-use of planar patch clamp chips

被引:10
作者
Kao, Liyo [1 ]
Abuladze, Natalia [1 ]
Shao, Xuesi M. [2 ]
McKeegan, Kevin [3 ]
Kurtz, Ira [1 ,4 ]
机构
[1] Univ Calif Los Angeles, David Geffen Sch Med, Dept Med, Los Angeles, CA 90095 USA
[2] Univ Calif Los Angeles, David Geffen Sch Med, Dept Neurobiol, Los Angeles, CA 90095 USA
[3] Univ Calif Los Angeles, David Geffen Sch Med, Dept Earth & Space Sci, Los Angeles, CA 90095 USA
[4] Univ Calif Los Angeles, David Geffen Sch Med, Brain Res Inst, Los Angeles, CA 90095 USA
关键词
Patch clamp; Planar chips; Electrophysiology; NA+-HCO3-COTRANSPORTER; MUSCLE-FIBERS; CHANNEL; CELLS; ION;
D O I
10.1016/j.jneumeth.2012.05.002
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The patch clamp technique is widely used for recording the activity of ion channels in single cells and lipid bilayers. Most platforms utilize borosilicate glass configured as a pipette, however more recently planar patch clamp chips have been developed that require less technical expertise. Planar patch clamp chips in systems like the Nanion Port-a-Patch are useful in that they allow more rapid throughput in drug screening studies. This technique also has the ability to perform rapid solution changes from the intracellular side. A current drawback with the planar patch clamp chips is the need to utilize a separate chip for each experiment. This increases the cost of each experiment and is due to the fact that the similar to 1 mu m aperture used for cell attachment is thought to retain cellular debris thereby preventing subsequent cell attachment and formation of G Omega seals. In the present study we have for the first time solved the technical problem of developing a simple protocol for re-use of Nanion planar patch clamp chips. The re-use methodology is demonstrated in whole cell patch clamp studies of HEK-293 cells expressing the electrogenic sodium bicarbonate cotransporter NBCe1-A in protocols involving external and internal solution changes, and CHO-K1 cells with incorporated gramicidin channels. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:205 / 210
页数:6
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