Müller D J, Engel A
M. E. Muller Institute for Microscopic Structural Biology, Biozentrum, University of Basel, Switzerland.
Biophys J. 1997 Sep;73(3):1633-44. doi: 10.1016/S0006-3495(97)78195-5.
In biological applications of atomic force microscopy, the different surface properties of the biological sample and its support become apparent. Observed height differences between the biomolecule and its supporting surface are thus not only of structural origin, but also depend on the different sample-tip and support-tip interactions. This can result in negative or positive contributions to the measured height, effects that are described by the DLVO (Derjaguin, Landau, Verwey, Overbeek) theory. Experimental verification shows that the electrostatic interactions between tip and sample can strongly influence the result obtained. To overcome this problem, pH and electrolyte concentration of the buffer solution have to be adjusted to screen out electrostatic forces. Under these conditions, the tip comes into direct contact with the surface of support and biological system, even when low forces required to prevent sample deformation are applied. In this case, the measured height can be related to the thickness of the native biological structure. The observed height dependence of the macromolecules on electrolyte concentration makes it possible to estimate surface charge densities.
在原子力显微镜的生物学应用中,生物样品及其支撑物的不同表面特性变得明显。因此,生物分子与其支撑表面之间观察到的高度差异不仅源于结构,还取决于不同的样品 - 探针和支撑 - 探针相互作用。这可能会对测量高度产生负或正的贡献,这些效应由DLVO(德亚金、朗道、韦韦、奥弗比克)理论描述。实验验证表明,探针与样品之间的静电相互作用会强烈影响所得结果。为克服此问题,必须调整缓冲溶液的pH值和电解质浓度以屏蔽静电力。在这些条件下,即使施加防止样品变形所需的低力,探针也会直接接触支撑物和生物系统的表面。在这种情况下,测量高度可与天然生物结构的厚度相关。观察到的大分子高度对电解质浓度的依赖性使得估计表面电荷密度成为可能。