Welcome to our book review site go-pdf.online!
You may have to Search all our reviewed books and magazines, click the sign up button below to create a free account.
Thermal Fluctuations of the Lipid Membrane Determine Particle Uptake Into Giant Unilamellar Vesicles
Abstract: Phagocytic particle uptake is crucial for the fate of both living cells and pathogens. Invading particles have to overcome fluctuating lipid membranes as the first physical barrier. However, the energy and the role of the fluctuation-based particle-membrane interactions during particle uptake are not understood. We tackle this problem by indenting the membrane of differently composed Giant Unilamellar Vesicles (GUVs) with optically trapped particles until particle uptake. By continuous 1 MHz tracking and autocorrelating the particle's positions within 30μs delays for different indentations, the fluctuations' amplitude, the damping, the mean forces, and the energy profiles were obtained. Remarkably, the uptake energy into a GUV becomes predictable since it increases for smaller fluctuation amplitudes and longer relaxation time. Our observations could be explained by a mathematical model based on continuous suppression of fluctuation modes. Hence, the reduced particle uptake energy for protein-ligand interactions LecA-Gb3 or Biotin-Streptavidin results also from pronounced, low-friction membrane fluctuations
A Shape-switch-block Method for Confocal Light-sheet Microscopy with Sectioned Bessel Beams and Stimulated Emission Depletion
Abstract: Microscopy seeks to simultaneously maximize optical resolution, contrast, speed, volume size, and probe tolerability, which is possible by combining different complementary imaging techniques with their specific strengths. Here, we show how to combine three physical concepts to increase resolution and contrast in light-sheet microscopy by making the effective light-sheet thinner through phase shaping, fluorophores-switching, and dynamic blocking of fluorescence. This shape-switch-block principle is realized by illumination with two holographically shaped, sectioned Bessel beams. Second, by switching off fluorophores in the proximity of the excitation center using continuous-wave stimulated emission depletion (STED). And third, by blocking fluorescence outside the switching region by confocal line detection. Thereby, we reduce the light-sheet thickness by 35%, achieving an isotropic resolution with beads in a 300 × 70 × 50 μm3 volume. Without STED, we obtain 0.37 μm resolution in cell clusters at improved sectioning and penetration depth. The shape-switch-block concept promises high potential, also for other microscopy techniques
ROCS Microscopy with Distinct Zero-order Blocking
Abstract: Research in modern light microscopy continuously seeks to improve spatial and temporal resolution in combination with user-friendly, cost-effective imaging systems. Among different label-free imaging approaches, Rotating Coherent Scattering (ROCS) microscopy in darkfield mode achieves superior resolution and contrast without image reconstructions, which is especially helpful in life cell experiments. Here we demonstrate how to achieve 145 nm resolution with an amplitude transmission mask for spatial filtering. This mask blocks the reflected 0-th order focus at 12 distinct positions, thereby increasing the effective aperture for the light back-scattered from the object. We further show how angular correlation analysis between coherent raw images helps to estimate the information content from different illumination directions
Super-resolution Microscopy and Single-molecule Tracking Reveal Distinct Adaptive Dynamics of MreB and of Cell Wall-synthesis Enzymes
Abstract: The movement of filamentous, actin-like MreB and of enzymes synthesizing the bacterial cell wall has been proposed to be highly coordinated. We have investigated the motion of MreB and of RodA and PbpH cell wall synthesis enzymes at 500 ms and at 20 ms time scales, allowing us to compare the motion of entire MreB filaments as well as of single molecules with that of the two synthesis proteins. While all three proteins formed assemblies that move with very similar trajectory orientation and with similar velocities, their trajectory lengths differed considerably, with PbpH showing shortest and MreB longest trajectories. These experiments suggest different on/off rates for RodA and Pb...
