The adsorption of DNA molecules onto a flat mica surface is a necessary step to perform atomic force microscopy studies of DNA conformation and observe DNA-protein interactions in physiological environment. However, the phenomenon that pulls DNA molecules onto the surface is still not understood. This is a crucial issue because the DNA/surface interactions could affect the DNA biological functions. In this paper we develop a model that can explain the mechanism of the DNA adsorption onto mica. This model suggests that DNA attraction is due to the sharing of the DNA and mica counterions. The correlations between divalent counterions on both the negatively charged DNA and the mica surface can generate a net attraction force whereas the correlations between monovalent counterions are ineffective in the DNA attraction. DNA binding is then dependent on the fractional surface densities of the divalent and monovalent cations, which can compete for the mica surface and DNA neutralizations. In addition, the attraction can be enhanced when the mica has been pretreated by transition metal cations (Ni(2+), Zn(2+)). Mica pretreatment simultaneously enhances the DNA attraction and reduces the repulsive contribution due to the electrical double-layer force. We also perform end-to-end distance measurement of DNA chains to study the binding strength. The DNA binding strength appears to be constant for a fixed fractional surface density of the divalent cations at low ionic strength (I < 0.1 M) as predicted by the model. However, at higher ionic strength, the binding is weakened by the screening effect of the ions. Then, some equations were derived to describe the binding of a polyelectrolyte onto a charged surface. The electrostatic attraction due to the sharing of counterions is particularly effective if the polyelectrolyte and the surface have nearly the same surface charge density. This characteristic of the attraction force can explain the success of mica for performing single DNA molecule observation by AFM. In addition, we explain how a reversible binding of the DNA molecules can be obtained with a pretreated mica surface.
Dispersion forces are present everywhere. Their importance, however, is largely neglected because
directly at a surface or at an interface they are mostly weak compared with specific interaction of short
range. Here, we show that these forces are nonetheless extremely relevant and may have drastic consequences
on the stability of thin films. We demonstrate that a force (per unit area) of <1 Pa is capable of “destroying”
100 nm (!) thick films, even if they are “glued” to the substrate by end-grafted polymers. We present the
temporal evolution of different morphologies of unstable thin liquid polymer films caused by destabilizing
intermolecular forces.
Adsorption of DNA molecules on mica, a highly negatively charged surface, mediated by divalent or trivalent cations is considered. By analyzing atomic force microscope (AFM) images of DNA molecules adsorbed on mica, phase diagrams of DNA molecules interacting with a mica surface are established in terms of concentrations of monovalent salt (NaCl) and divalent (MgCl2) or multivalent (spermidine, cobalt hexamine) salts. These diagrams show two transitions between nonadsorption and adsorption. The first one arises when the concentration of multivalent counterions is larger than a limit value, which is not sensitive to the monovalent salt concentration. The second transition is due to the binding competition between monovalent and multivalent counterions. In addition, we develop a model of polyelectrolyte adsorption on like-charged surfaces with multivalent counterions. This model shows that the correlations of the multivalent counterions at the interface between DNA and mica play a critical role. Furthermore, it appears that DNA adsorption takes place when the energy gain in counterion correlations overcomes an energy barrier. This barrier is induced by the entropy loss in confining DNA in a thin adsorbed layer, the entropy loss in the interpenetration of the clouds of mica and DNA counterions, and the electrostatic repulsion between DNA and mica. The analysis of the experimental results provides an estimation of this energy barrier. We then discuss some important issues, including DNA adsorption under physiological conditions.
Long-range van der Waals forces may govern the physical behavior of thin films with thicknesses lying
in the nanometer range scale. Here, we show that these forces can also be highly efficient across multilayer
systems. The observed destabilization of a thin fluid layer, confined between a rigid substrate and a thin
solid polymeric film, leading to the deformation of the solid covering film, revealed the strength of these
interactions. Furthermore, simply by modifying the thickness of the layers or by changing the bounding
medium, the effect of these interactions could be controlled and even reversed.
We use optical and scanning force microscopy to explore the possibility of switching the stability of a bilayer of poly(methyl methacrylate) (PMMA) on polystyrene by simply changing the film thickness. We show that for thin PMMA layers on thicker polystyrene films the PMMA layer is unstable to thickness fluctuations. However, polystyrene layers are unstable when they are substantially thinner than the now stable PMMA film. Dewetting morphologies are cataloged as a function of the thickness of individual polymer layers by identifying which layer is unstable by which mechanism, be it spinodal dewetting or heterogeneous thermal nucleation. Our results are in good agreement with a linear stability analysis of the influence of long-range dispersion forces, but also indicate the influence of film preparation and small variations of material properties.
The atomic force microscope is a key tool for investigating DNA conformation and DNA-protein interactions in liquid. The main advantage of this technique is that moving molecules can be studied in real time provided that molecules are sufficiently bound to the surface. Mg 2+ ions with a very low concentration of monovalent salt are generally used to attach DNA on mica because monovalent counterions inhibit the DNA electrostatic attraction with the surface. However, monovalent counterions at physiological concentrations are necessary to obtain specific DNA/protein interactions. To solve this problem, we propose a new protocol to obtain a reversible binding of DNA on NiCl 2-pretreated mica. This protocol uses Mg 2+ ions for monitoring DNA attachment on NiCl2-pretreated mica, which allows the DNA molecules to remain bound to the surface even at high NaCl concentration thanks to Ni 2+ ions adsorbed on the surface.
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