An official website of the United States government
There is considerable interest in measuring, with nanoscale spatial resolution, the physical properties of lipid membranes because of their role in the physiology of living systems. Due to its ability to nondestructively image surfaces in solution, tapping mode atomic force microscopy (TMAFM) has proven to be a useful technique for imaging lipid membranes. However, further information concerning the mechanical properties of surfaces is contained within the time-resolved tip/sample force interactions. The tapping forces can be recovered by taking the second derivative of the cantilever deflection signal and scaling by the effective mass of the cantilever; this technique is referred to as scanning probe acceleration microscopy. Herein, we describe how the maximum and minimum tapping forces change with surface mechanical properties. Furthermore, we demonstrate how these changes can be used to measure mechanical changes in lipid membranes containing cholesterol.
more »
« less
Investigation of Protein/Lipid Interactions via Scanning Probe Acceleration Microscopy: Theory and Experiment
There is great interest in the application of proximal probe techniques to simultaneously image and measure mechancial properties of surfaces with nanoscale spatial resolution. There have been several innovations in generating time-resolved force interaction between the tip and surface while acquiring a tapping mode AFM image. These tip/sample forces contain information regarding mechanical properties of surfaces in an analogous fashion to a force curve experiment. Here, we demonstrate, via simulation, that the maximum and minimum tapping forces change with respect to the Young’s modulus and adhesiveness of a surface, but the roughness of the surfaces has no effect on the tapping forces. Using these changes in tapping forces, we determine the mechanical changes of a lipid membrane after exposure to a huntingtin exon1 (htt exon1) protein with an expanded polyglutamine (polyQ) domain. Expanded polyQ domains in htt is associated with Huntington’s disease, a genetic neurodegenerative disorder. The htt exon1 protein caused regions of increased surface roughness to appear in the lipid membrane, and these areas were associated with decreased elasticity and adhesion to the AFM probe.
more »
« less
- Award ID(s):
- 1054211
- PAR ID:
- 10018320
- Date Published:
- Journal Name:
- ASME 2012 International Design Engineering Technical Conferences and Computers and Information in Engineering Conference
- Volume:
- 5
- Page Range / eLocation ID:
- 297-302
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
Precisely quantifying the energetics that drive the folding of membrane proteins into a lipid bilayer remains challenging. More than 15 years ago, atomic force microscopy (AFM) emerged as a powerful tool to mechanically extract individual membrane proteins from a lipid bilayer. Concurrently, fluctuation theorems, such as the Jarzynski equality, were applied to deduce equilibrium free energies (ΔG0) from non-equilibrium single-molecule force spectroscopy records. The combination of these two advances in single-molecule studies deduced the free-energy of the model membrane protein bacteriorhodopsin in its native lipid bilayer. To elucidate this free-energy landscape at a higher resolution, we applied two recent developments. First, as an input to the reconstruction, we used force-extension curves acquired with a 100-fold higher time resolution and 10-fold higher force precision than traditional AFM studies of membrane proteins. Next, by using an inverse Weierstrass transform and the Jarzynski equality, we removed the free energy associated with the force probe and determined the molecular free-energy landscape of the molecule under study, bacteriorhodopsin. The resulting landscape yielded an average unfolding free energy per amino acid (aa) of 1.0 ± 0.1 kcal/mol, in agreement with past single-molecule studies. Moreover, on a smaller spatial scale, this high-resolution landscape also agreed with an equilibrium measurement of a particular three-aa transition in bacteriorhodopsin that yielded 2.7 kcal/mol/aa, an unexpectedly high value. Hence, while average unfolding ΔG0 per aa is a useful metric, the derived high-resolution landscape details significant local variation from the mean. More generally, we demonstrated that, as anticipated, the inverse Weierstrass transform is an efficient means to reconstruct free-energy landscapes from AFM data.more » « less
-
Determination of the surface hydrophobicity or wettability of nanomaterials and nanoparticles (NPs) is often challenged by the heterogeneous properties of NPs that vary with particle size, shape, surface charge, aggregation states, and surface sorption or coating. This study first summarized inherent limitations of the water contact angle, octanol–water partition coefficient ( K ow ) and surface adsorption of probe molecules in probing nanomaterial hydrophobicity. Then, we demonstrated the principle of a scanning probe method based on atomic force microscopy (AFM) for the local surface hydrophobicity measurement. Specifically, we measured the adhesion forces between functionalized AFM tips and self-assembled monolayers (SAMs) to establish a linear relationship between the adhesion forces and water contact angles based on the continuum thermodynamic approach (CTA). This relationship was used to determine the local surface hydrophobicity of seven different NPs ( i.e. , TiO 2 , ZnO, SiO 2 , CuO, CeO 2 , α-Fe 2 O 3 , and Ag), which agreed well with bulk contact angles of these NPs. Some discrepancies were observed for Fe 2 O 3 , CeO 2 and SiO 2 NPs, probably because of surface hydration and roughness effects. Moreover, the solution pH and ionic strength had negligible effects on the adhesion forces between the AFM tip and MWCNTs or C 60 , indicating that the hydrophobicity of carbonaceous nanomaterials is not influenced by pH or ionic strength (IS). By contrast, natural organic matter (NOM) appreciably decreased the hydrophobicity of MWCNTs and C 60 due to surface coating of hydrophilic NOM. This scanning probe method has been proved to be reliable and robust toward the accurate measurement of the nanoscale hydrophobicity of individual NPs or nanomaterials in liquid environments.more » « less
-
Historically, high carbon steels have been used in mechanical applications because their high surface hardness contributes to excellent wear performance. However, in aggressive environments, current bearing steels exhibit insufficient corrosion resistance. Martensitic stainless steels are attractive for bearing applications due to their high corrosion resistance and ability to be surface hardened via carburizing heat treatments. Here three different carburizing heat treatments were applied to UNS S42670: a high-temperature temper (HTT), a low-temperature temper (LTT), and carbo-nitriding (CN). Magnetic force microscopy showed differences in magnetic domains between the matrix and carbides, while scanning Kelvin probe force microscopy (SKPFM) revealed a 90–200 mV Volta potential difference between the two phases. Corrosion progression was monitored on the nanoscale via SKPFM and in situ atomic force microscopy (AFM), revealing different corrosion modes among heat treatments that predicted bulk corrosion behavior in electrochemical testing. HTT outperforms LTT and CN in wear testing and thus is recommended for non-corrosive aerospace applications, whereas CN is recommended for corrosion-prone applications as it exhibits exceptional corrosion resistance. The results reported here support the use of scanning probe microscopy for predicting bulk corrosion behavior by measuring nanoscale surface differences in properties between carbides and the surrounding matrix.more » « less
-
Johnson, Colin (Ed.)Lipid membranes in nature adapt and reconfigure to changes in composition, temperature, humidity, and mechanics. For instance, the oscillating mechanical forces on lung cells and alveoli influence membrane synthesis and structure during breathing. However, despite advances in the understanding of lipid membrane phase behavior and mechanics of tissue, there is a critical knowledge gap regarding the response of lipid membranes to micromechanical forces. Most studies of lipid membrane mechanics use supported lipid bilayer systems missing the structural complexity of pulmonary lipids in alveolar membranes comprising multi-bilayer interconnected stacks. Here, we elucidate the collective response of the major component of pulmonary lipids to strain in the form of multi-bilayer stacks supported on flexible elastomer substrates. We utilize X-ray diffraction, scanning probe microscopy, confocal microscopy, and molecular dynamics simulation to show that lipid multilayered films both in gel and fluid states evolve structurally and mechanically in response to compression at multiple length scales. Specifically, compression leads to increased disorder of lipid alkyl chains comparable to the effect of cholesterol on gel phases as a direct result of the formation of nanoscale undulations in the lipid multilayers, also inducing buckling delamination and enhancing multi-bilayer alignment. We propose this cooperative short- and long-range reconfiguration of lipid multilayered films under compression constitutes a mechanism to accommodate stress and substrate topography. Our work raises fundamental insights regarding the adaptability of complex lipid membranes to mechanical stimuli. This is critical to several technologies requiring mechanically reconfigurable surfaces such as the development of electronic devices interfacing biological materials.more » « less
- Free Publicly Accessible Full Text
-
Accepted Manuscript1.0
- Conference Paper:
- https://doi.org/10.1115/DETC2012-70228
