Investigation of temperature induced mechanical changes in supported bilayers by variants of tapping mode atomic...

Tapping mode atomic force microscopy (AFM) is an invaluable technique for examining topographical features of biological materials in solution, and there has been a growing interest in developing techniques to provide further compositional contrast and information concerning surface mechanical properties. Phase shifts, cantilever response at higher harmonic frequencies of the drive, and time-resolved tip/sample force reconstruction have all been shown to provide additional compositional contrast of surfaces, as compared to basic tapping mode AFM imaging. This study aimed to demonstrate the relative ability of these different imaging techniques to detect temperature induced changes in the elastic modulus of supported total brain lipid extract (TBLE) bilayer patches on mica. To aid in direct comparison between the different imaging techniques, all required data was obtained simultaneously while capturing traditional tapping mode AFM topography images. While all of the techniques were able to provide compositional contrast consistent with known temperature-induced changes in the bilayer patch, interpretation of the resulting contrast was not always straightforward. Phase imaging suffered from contrast inversion. Individual harmonics responded in a variety of ways to the temperature-induced changes in elastic modulus of the bilayer. The maximum tapping force (or peak force) associated with imaging the bilayer correctly reflected the changes in elastic modulus of the lipid bilayer. Importantly, as the required data can be obtained simultaneously, combining these different imaging techniques can lead to a more complete understanding of a sample's mechanical features.