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note
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In the constant-deflection mode, a feedback loop is used to keep the normal deflection of the cantilever constant. The force applied to the surface is held constant in this imaging mode.
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Adhesive force maps were obtained by recording 64 × 64 force curves per image. From this force curve arrays, x-y adhesion maps were generated by displaying the pull-off force measured for each force curve. Images were resampled to 512 × 512 pixels.
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48
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5244279829
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note
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The height differences were measured from cross sections in the topographic images. At forces ∼1 nN, due to variations in the material properties of the two lipids, the step height varied from one probe to another as a result of variations in the radius of curvature of the probe. We present a mean value and standard deviation of height differences from six different images.
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The monolayer was cut by imaging 1 μm × 1 μm areas at large forces (>1500 nN) and high rates (60 Hz) for short period of times. Larger images of these areas under normal loads with new probes revealed terraces with 1 nm step heights which is consistent with half the unit cell of the muscovite mica lattice along the c axis (Hu, J.; Xiao, X.-d.; Ogletree, D. F.; Salmeron, M. Surf. Sci. 1995, 327, 358). Complete removal of the lipid film allowed us to unambiguously measure the film thickness.
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note
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In the presence of attractive forces such as observed in this study, the effective force applied on the surface is equivalent to the sum of the externally applied force and of the adhesive pull-off force.
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52
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5244381258
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note
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3 gives a monolayer thickness of 3.0 nm. Note that such calculation cannot be done accurately for DOPE due to its unknown density.
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54
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5244305425
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note
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Mean value and standard deviation of 10 pull-off forces measured in three different regions. The absolute values quoted for the adhesive forces can vary from one experiment to another due to changes in probe chemistry and geometry.
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note
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At forces ∼1 nN, images of the same area could be obtained without altering the sample. At large imaging forces (>5 nN) a decreased topographic contrast could sometimes be observed. Various scan speeds were tested (0.5-10 Hz, corresponding to 15-300 μm/s) and were found to have no influence on the topographic and friction contrasts. Trace and retrace images were inverted for friction and identical for topography, indicating no significant contribution of lateral forces to the apparent topographic contrast.
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note
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An accurate estimate of the load exerted on the hydrated DSPE surface at the snap to contact would require a knowledge of the "mechanical properties" of the hydration layer. Lacking this information we can estimate a lower limit of pressure of ∼3 MPa, by assuming that the 4 nN force is distributed over the entire probe area (∼20 nm radius). The actual pressure exerted by the apex of the probe is much larger than this lower limit.
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Note that two modes of imaging based on repulsive interactions have been recently reported. Senden, T. J.; Drummond, C. J.; Kékicheff, P. Langmuir 1994, 10, 358. Manne, S.; Cleveland, J. P.; Gaub, H. E.; Stucky, G. D.; Hansma, P. K. Langmuir 1994, 10, 4409.
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