CEA, DSV, iRTSV/Biopuces, Grenoble Cedex 9, France.
Biomaterials. 2010 Oct;31(28):7398-410. doi: 10.1016/j.biomaterials.2010.06.015.
Planar patch-clamp is a two-dimensional variation of traditional patch-clamp. By contrast to classical glass micropipette, the seal quality of silicon patch-clamp chips (i.e. seal resistance and seal success rate) have remained poor due to the planar geometry and the nature of the substrate and thus partially obliterate the advantages related to planar patch-clamp. The characterization of physical parameters involved in seal formation is thus of major interest. In this paper, we demonstrate that the physical characterization of surfaces by a set of techniques (Atomic Force Microscopy (AFM), Scanning Electron Microscopy (SEM), X-ray Photoelectron Spectroscopy (XPS), surface energy (polar and dispersive contributions), drop angles, impedance spectroscopy, combined with a statistical design of experiments (DOE)) allowed us discriminating chips that provide relevant performances for planar patch-clamp analysis. Analyses of seal quality demonstrate that dispersive interactions and micropore size are the most crucial physical parameters of chip surfaces, by contrast to surface roughness and dielectric membrane thickness. This multi-scale study combined with electrophysiological validation of chips on a diverse set of cell-types expressing various ion channels (IRK1, hERG and hNa(v)1.5 channels) unveiled a suitable patch-clamp chip candidate. This original approach may inspire novel strategies for selecting appropriate surface parameters dedicated to biochips.
平面片层钳位是传统片层钳位的二维变体。与经典的玻璃微量移液器相比,由于平面几何形状和基底的性质,硅片层钳位芯片的密封质量(即密封电阻和密封成功率)仍然很差,从而部分抹杀了与平面片层钳位相关的优势。因此,与密封形成相关的物理参数的特性描述是主要关注点。在本文中,我们证明了通过一组技术(原子力显微镜(AFM)、扫描电子显微镜(SEM)、X 射线光电子能谱(XPS)、表面能(极性和分散贡献)、滴角、阻抗谱)对表面进行物理特性描述,结合实验设计(DOE)的统计分析,我们能够区分出那些能够为平面片层钳位分析提供相关性能的芯片。通过对不同类型的表达各种离子通道(IRK1、hERG 和 hNa(v)1.5 通道)的细胞进行的密封质量分析表明,与表面粗糙度和介电膜厚度相比,分散相互作用和微孔尺寸是芯片表面最关键的物理参数。这项多尺度研究结合了对不同细胞类型的芯片进行电生理验证,揭示了一种合适的片层钳位芯片候选者。这种新颖的方法可能为选择专门用于生物芯片的合适表面参数提供新的策略。