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基于原子力显微镜和电化学阻抗谱技术在硅基微腔中研究脂双层膜。

Lipid Bilayer Membrane in a Silicon Based Micron Sized Cavity Accessed by Atomic Force Microscopy and Electrochemical Impedance Spectroscopy.

机构信息

Electrical and Computer Engineering Department, University of Alabama in Huntsville, Huntsville, AL 35899, USA.

Biotechnology Science and Engineering Program, University of Alabama in Huntsville, Huntsville, AL 35899, USA.

出版信息

Biosensors (Basel). 2017 Jul 5;7(3):26. doi: 10.3390/bios7030026.

Abstract

Supported lipid bilayers (SLBs) are widely used in biophysical research to probe the functionality of biological membranes and to provide diagnoses in high throughput drug screening. Formation of SLBs at below phase transition temperature () has applications in nano-medicine research where low temperature profiles are required. Herein, we report the successful production of SLBs at above-as well as below-the of the lipids in an anisotropically etched, silicon-based micro-cavity. The Si-based cavity walls exhibit controlled temperature which assist in the quick and stable formation of lipid bilayer membranes. Fusion of large unilamellar vesicles was monitored in real time in an aqueous environment inside the Si cavity using atomic force microscopy (AFM), and the lateral organization of the lipid molecules was characterized until the formation of the SLBs. The stability of SLBs produced was also characterized by recording the electrical resistance and the capacitance using electrochemical impedance spectroscopy (EIS). Analysis was done in the frequency regime of 10-10⁵ Hz at a signal voltage of 100 mV and giga-ohm sealed impedance was obtained continuously over four days. Finally, the cantilever tip in AFM was utilized to estimate the bilayer thickness and to calculate the rupture force at the interface of the tip and the SLB. We anticipate that a silicon-based, micron-sized cavity has the potential to produce highly-stable SLBs below their . The membranes inside the Si cavity could last for several days and allow robust characterization using AFM or EIS. This could be an excellent platform for nanomedicine experiments that require low operating temperatures.

摘要

支持性脂质双层 (SLB) 在生物物理研究中被广泛用于探测生物膜的功能,并在高通量药物筛选中提供诊断。低于相变温度 ( ) 下形成 SLB 在需要低温剖面的纳米医学研究中有应用。在此,我们报告了在各向异性蚀刻的基于硅的微腔中成功地在高于和低于脂质的 下制备 SLB。基于硅的腔壁具有受控的温度,这有助于快速且稳定地形成脂质双层膜。使用原子力显微镜 (AFM) 在 Si 腔内部的水相中实时监测大单层囊泡的融合,并对脂质分子的横向组织进行表征,直到形成 SLB。还使用电化学阻抗谱 (EIS) 来表征所制备的 SLB 的稳定性。在 10-10⁵ Hz 的频率范围内进行分析,信号电压为 100 mV,连续四天获得千兆欧姆密封阻抗。最后,在 AFM 中的悬臂尖端用于估计双层厚度,并计算尖端和 SLB 界面处的破裂力。我们预计,基于硅的微米级腔具有在低于其 下产生高度稳定的 SLB 的潜力。Si 腔内部的膜可以持续数天,并允许使用 AFM 或 EIS 进行稳健的表征。这可能是需要低温操作的纳米医学实验的绝佳平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bf7/5618032/3c8a713f2474/biosensors-07-00026-g001.jpg

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