Cell to aperture interaction in patch‐clamp chips visualized by fluorescence microscopy and focused‐ion beam sections
Cell to aperture interaction in patch‐clamp chips visualized by fluorescence microscopy and focused‐ion beam sections
复制标题
通过荧光显微镜和聚焦离子束切片可视化膜片钳芯片中细胞与孔径的相互作用
DOI：
10.1002/bit.23127
复制
发表时间：
2011
影响因子：
3.8
通讯作者：
G. Mealing
中科院分区：
文献类型：
--
作者：
C. Py;D. Jayus Nor Salim;R. Monette;T. Comas;J. Fraser;Dolores Martinez;Marzia Martina;G. Mealing
膜片钳是监测细胞电生理活动和药理化合物对特定离子通道蛋白作用的重要方法。近年来，平面贴片钳芯片作为一种高通量的方法被开发出来，以取代现有的玻璃移液法。然而，适当的条件，以优化高电阻电池-探针密封所需的测量小电流产生的离子通道活性仍然是猜测的主题。在这里，我们报道了多孔径（筛）芯片的设计，以快速方便地评估细胞与孔径之间的相互作用。我们提出了一种基于染料加载协议的方法，通过芯片的孔径和随后的荧光共聚焦显微镜评估来预筛选密封的质量。我们还展示了在贴片钳芯片孔径中细胞聚焦离子束截面的第一张扫描电子显微照片。Biotechnol。Bioeng。2011;108:1936 - 1941。©2011 Wiley期刊公司
Patch‐clamp is an important method to monitor the electrophysiological activity of cells and the role of pharmacological compounds on specific ion channel proteins. In recent years, planar patch‐clamp chips have been developed as a higher throughput approach to the established glass‐pipette method. However, proper conditions to optimize the high resistance cell‐to‐probe seals required to measure the small currents resulting from ion channel activity are still the subject of conjecture. Here, we report on the design of multiple‐aperture (sieve) chips to rapidly facilitate assessment of cell‐to‐aperture interactions in statistically significant numbers. We propose a method to pre‐screen the quality of seals based on a dye loading protocol through apertures in the chip and subsequent evaluation with fluorescence confocal microscopy. We also show the first scanning electron micrograph of a focused ion beam section of a cell in a patch‐clamp chip aperture. Biotechnol. Bioeng. 2011; 108:1936–1941. © 2011 Wiley Periodicals, Inc.