Goyal Gaurav, Darvish Armin, Kim Min Jun
School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, PA 19104, USA.
Analyst. 2015 Jul 21;140(14):4865-73. doi: 10.1039/c5an00250h. Epub 2015 Mar 26.
Membrane deformation of nano-vesicles is crucial in many cellular processes such as virus entry into the host cell, membrane fusion, and endo- and exocytosis; however, studying the deformation of sub-100 nm soft vesicles is very challenging using the conventional techniques. In this paper, we report detecting co-translocational deformation of individual 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) nano-liposomes using solid-state nanopores. Electrokinetic translocation through the nanopore caused the soft DOPC liposomes (85 nm diameter) to change their shape, which we attribute to the strong electric field strength and physical confinement inside the pore. The experiments were performed at varying transmembrane voltages and the deformation was observed to mount up with increasing applied voltage and followed an exponential trend. Numerical simulations were performed to simulate the concentrated electric field strength inside the nanopore and a field strength of 14 kV cm(-1) (at 600 mV applied voltage) was achieved at the pore center. The electric field strength inside the nanopore is much higher than the field strength known to cause deformation of 15-30 μm giant membrane vesicles. As a control, we also performed experiments with rigid polystyrene beads that did not show any deformation during translocation events, which further established our hypothesis of co-translocational deformation of liposomes. Our technique presents an innovative and high throughput means for investigating deformation behavior of soft nano-vesicles.
纳米囊泡的膜变形在许多细胞过程中至关重要,如病毒进入宿主细胞、膜融合以及胞吞和胞吐作用;然而,使用传统技术研究直径小于100 nm的软囊泡的变形极具挑战性。在本文中,我们报告了利用固态纳米孔检测单个1,2 - 二油酰 - sn - 甘油 - 3 - 磷酸胆碱(DOPC)纳米脂质体的共转运变形。通过纳米孔的电动转运导致柔软的DOPC脂质体(直径85 nm)改变其形状,我们将此归因于孔内的强电场强度和物理限制。实验在不同的跨膜电压下进行,观察到变形随着施加电压的增加而加剧,并呈指数趋势。进行了数值模拟以模拟纳米孔内的集中电场强度,在孔中心实现了14 kV cm⁻¹(在600 mV施加电压下)的场强。纳米孔内的电场强度远高于已知会导致直径15 - 30 μm的巨型膜囊泡变形的场强。作为对照,我们还用刚性聚苯乙烯珠进行了实验,这些珠子在转运过程中未显示任何变形,这进一步证实了我们关于脂质体共转运变形的假设。我们的技术为研究软纳米囊泡的变形行为提供了一种创新且高通量的方法。