Batishchev Oleg V, Indenbom Andrey V
A.N.Frumkin Institute of Physical Chemistry and Electrochemistry RAS, 31 Leninsky prosp., 119991 Moscow, Russia.
Bioelectrochemistry. 2008 Nov;74(1):22-5. doi: 10.1016/j.bioelechem.2008.02.002. Epub 2008 Feb 29.
Formation of bilayer lipid membrane (BLM) by Montal-Mueller technique across a small aperture in a partition film traditionally requires coating of the aperture with a hydrophobic substance, often just an organic solvent. However, we demonstrate here that the most effective coating is not strictly hydrophobic but rather provides water/oil repellent properties. BLM were formed from diphytanoylphosphatidylcholine (DPhPC) on small 0.1-0.8 mm apertures made in specially prepared alkylated glass coverslips. The coverslips were either fluorosiliconized by 3,3,3-Trifluoropropyl-trimethoxysilane, which reduces adsorption of DPhPC in addition to creation of hydrophobic surface, or silanized, which promote adsorption of DPhPC. At fluorosiliconized surfaces stable BLM were formed. Specific capacitance of these BLM was 0.86 microF/cm(2)+/-5%, while their lateral tension was estimated as 4.3+/-0.4 mN/m. BLM were stable for hours under moderate voltage applied. At silanized surfaces stable BLM were formed only in acidic medium (3 <pH <4), while at higher pH the membranes could cover the aperture only partially. Thus, apertures in fluorosiliconized glass can be robustly used for formation of model lipid membranes under physiological conditions.
传统上,采用蒙塔尔-米勒技术在分隔膜的小孔上形成双层脂质膜(BLM)需要用疏水性物质(通常只是一种有机溶剂)涂覆小孔。然而,我们在此证明,最有效的涂层并非严格意义上的疏水性,而是具有拒水/拒油性能。BLM由二植烷酰磷脂酰胆碱(DPhPC)在特制的烷基化玻璃盖玻片上制作的0.1 - 0.8毫米的小孔上形成。盖玻片要么用3,3,3 - 三氟丙基三甲氧基硅烷进行氟硅化处理,这除了能形成疏水表面外,还能减少DPhPC的吸附,要么进行硅烷化处理,这会促进DPhPC的吸附。在氟硅化表面形成了稳定的BLM。这些BLM的比电容为0.86微法/平方厘米±5%,而其侧向张力估计为4.3±0.4毫牛/米。在施加适度电压的情况下,BLM能稳定数小时。在硅烷化表面,仅在酸性介质(3<pH<4)中形成了稳定的BLM,而在较高pH值时,膜只能部分覆盖小孔。因此,氟硅化玻璃中的小孔可在生理条件下稳健地用于形成模型脂质膜。