AFM was proven suitable for determining the stiffness of ONH tissues, encouraging further large-scale analyses. The marked decrease in stiffness, implying an increased deformability of the ONH in PEX eyes, may reflect an inherent tissue weakness rendering these eyes more vulnerable to glaucomatous damage.
SiiE from Salmonella enterica is a giant 5,559-residue-long nonfimbrial adhesin that is secreted by a type 1 secretion system (T1SS) and initiates bacterial adhesion to polarized host cells. Structural insight has been gained into the 53 bacterial Ig-like (BIg) domains of SiiE, which account for 94% of the entire SiiE sequence. The crystal structure of a fragment comprising BIg domains 50 to 52 of SiiE reveals the BIg domain architecture and highlights two types of SiiE-specific Ca²⁺-binding sites. Sequence homology considerations suggest that full-length SiiE interacts with more than 100 Ca²⁺ ions. Molecular dynamics simulations and single-molecule imaging indicate that Ca²⁺ binding confers SiiE with a rigid 200 nm rod-like habitus that is required to reach out beyond the Salmonella lipopolysaccharide layer and to promote adhesion to host cells. The crystal structure suggests plausible routes for the establishment of the initial contact between Salmonella and host cells.
We present a high-speed atomic force microscope that exhibits a number of practical advantages over previous designs. Its central component is a high-speed scanner with a maximum scan size of 23 microm x 23 microm and a conveniently large sample stage area (6.5 mm x 6.5 mm). In combination with small cantilevers, image rates of up to 46 images s(-1) in air and 13 images s(-1) in liquid are reached under z-feedback control. By large scan size imaging of collagen fibrils in air, sample velocities of 8.8 mm s(-1) in the xy-direction and 11 mm s(-1) in the z-direction are reached. To provide optimized imaging conditions for both large and small scan sizes, a modular scanner design allows easily exchanging the x- and y-piezos. The scanner is therefore also suited for investigations on the molecular and atomic scale, which is demonstrated by imaging the step dynamics of a calcite surface during dissolution and the hexagonal lattice of a mica surface in liquid.
The imaging speed of the wide-spread force mapping mode for quantitative mechanical measurements on soft samples in liquid with the atomic force microscope (AFM) is limited by the bandwidth of the z-scanner and viscous drag forces on the cantilever. Here, we applied high-speed, large scan-range atomic force microscopy and small cantilevers to increase the speed of force mapping by ≈10-100 times. This allowed resolving dynamic processes on living mouse embryonic fibroblasts. Cytoskeleton reorganization during cell locomotion, growth of individual cytoskeleton fibers, cell blebbing, and the formation of endocytic pits in the cell membrane were observed. Increasing the force curve rate from 2 to 300 Hz increased the measured apparent Young's modulus of the cells by about 10 times, which facilitated force mapping measurements at high speed.
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