scholarly journals Mechanical heterogeneity of dentin at different length scales as determined by AFM phase contrast

Micron ◽  
2012 ◽  
Vol 43 (12) ◽  
pp. 1364-1371 ◽  
Author(s):  
Luiz E. Bertassoni ◽  
Grayson W. Marshall ◽  
Michael V. Swain
eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Robert J Huebner ◽  
Abdul Naseer Malmi-Kakkada ◽  
Sena Sarikaya ◽  
Shinuo Weng ◽  
D Thirumalai ◽  
...  

Morphogenesis is governed by the interplay of molecular signals and mechanical forces across multiple length scales. The last decade has seen tremendous advances in our understanding of the dynamics of protein localization and turnover at sub-cellular length scales, and at the other end of the spectrum, of mechanics at tissue-level length scales. Integrating the two remains a challenge, however, because we lack a detailed understanding of the subcellular patterns of mechanical properties of cells within tissues. Here, in the context of the elongating body axis of Xenopus embryos, we combine tools from cell biology and physics to demonstrate that individual cell-cell junctions display finely-patterned local mechanical heterogeneity along their length. We show that such local mechanical patterning is essential for the cell movements of convergent extension and is imparted by locally patterned clustering of a classical cadherin. Finally, the patterning of cadherins and thus local mechanics along cell-cell junctions are controlled by Planar Cell Polarity signaling, a key genetic module for CE that is mutated in diverse human birth defects.


Author(s):  
Robert J. Huebner ◽  
Abdul Naseer Malmi-Kakkada ◽  
Sena Sarikaya ◽  
Shinuo Weng ◽  
D. Thirumalai ◽  
...  

AbstractMorphogenesis is governed by the interplay of molecular signals and mechanical forces across multiple length scales. The last decade has seen tremendous advances in our understanding of the dynamics of protein localization and turnover at sub-cellular length scales, and at the other end of the spectrum, of mechanics at tissue-level length scales. Integrating the two remains a challenge, however, because we lack a detailed understanding of the subcellular patterns of mechanical properties of cells within tissues. Here, in the context of the elongating body axis of a vertebrate embryo, we combine tools from cell biology and physics to demonstrate that individual cell-cell junctions display finely-patterned local mechanical heterogeneity along their length. We show that such local mechanical patterning is essential for the cell movements of convergent extension and is imparted by locally patterned clustering of a classical cadherin. Finally, the patterning of cadherins and thus local mechanics along cell-cell junctions are controlled by Planar Cell Polarity signaling, a key genetic module for CE that is mutated in diverse human birth defects.


Author(s):  
Mitsuo Ohtsuki ◽  
Michael Sogard

Structural investigations of biological macromolecules commonly employ CTEM with negative staining techniques. Difficulties in valid image interpretation arise, however, due to problems such as variability in thickness and degree of penetration of the staining agent, noise from the supporting film, and artifacts from defocus phase contrast effects. In order to determine the effects of these variables on biological structure, as seen by the electron microscope, negative stained macromolecules of high density lipoprotein-3 (HDL3) from human serum were analyzed with both CTEM and STEM, and results were then compared with CTEM micrographs of freeze-etched HDL3. In addition, we altered the structure of this molecule by digesting away its phospholipid component with phospholipase A2 and look for consistent changes in structure.


Author(s):  
S. Golladay

The theory of multiple scattering has been worked out by Groves and comparisons have been made between predicted and observed signals for thick specimens observed in a STEM under conditions where phase contrast effects are unimportant. Independent measurements of the collection efficiencies of the two STEM detectors, calculations of the ratio σe/σi = R, where σe, σi are the total cross sections for elastic and inelastic scattering respectively, and a model of the unknown mass distribution are needed for these comparisons. In this paper an extension of this work will be described which allows the determination of the required efficiencies, R, and the unknown mass distribution from the data without additional measurements or models. Essential to the analysis is the fact that in a STEM two or more signal measurements can be made simultaneously at each image point.


Author(s):  
J.M. Cowley

The problem of "understandinq" electron microscope imaqes becomes more acute as the resolution is improved. The naive interpretation of an imaqe as representinq the projection of an atom density becomes less and less appropriate. We are increasinqly forced to face the complexities of coherent imaqinq of what are essentially phase objects. Most electron microscopists are now aware that, for very thin weakly scatterinq objects such as thin unstained bioloqical specimens, hiqh resolution imaqes are best obtained near the optimum defocus, as prescribed by Scherzer, where the phase contrast imaqe qives a qood representation of the projected potential, apart from a lack of information on the lower spatial frequencies. But phase contrast imaqinq is never simple except in idealized limitinq cases.


Author(s):  
Y. Harada ◽  
T. Goto ◽  
H. Koike ◽  
T. Someya

Since phase contrasts of STEM images, that is, Fresnel diffraction fringes or lattice images, manifest themselves in field emission scanning microscopy, the mechanism for image formation in the STEM mode has been investigated and compared with that in CTEM mode, resulting in the theory of reciprocity. It reveals that contrast in STEM images exhibits the same properties as contrast in CTEM images. However, it appears that the validity of the reciprocity theory, especially on the details of phase contrast, has not yet been fully proven by the experiments. In this work, we shall investigate the phase contrast images obtained in both the STEM and CTEM modes of a field emission microscope (100kV), and evaluate the validity of the reciprocity theory by comparing the experimental results.


Author(s):  
Juan Mora-Galindo ◽  
Jorge Arauz-Contreras

The zinc iodide-osmium tetroxide (ZIO) technique is presently employed to study both, neural and non neural tissues. Precipitates depends on cell types and possibly cell metabol ism as well.Guinea pig cecal mucosa, already known to be composed of epithelium with cells at different maturation stages and lamina propria which i s formed by morphologically and functionally heterogeneous cell population, was studied to determine the pat tern of ZIO impregnation. For this, adult Guinea pg cecal mucosa was fixed with buffered 1.2 5% g 1 utara 1 dehyde before incubation with ZIO for 16 hours, a t 4°C in the dark. Further steps involved a quick sample dehydration in graded ethanols, embedding in Epon 812 and sectioning to observe the unstained material under a phase contrast light microscope (LM) and a transmission electron microscope (TEM).


Author(s):  
N.V. Belov ◽  
U.I. Papiashwili ◽  
B.E. Yudovich

It has been almost universally adopted that dissolution of solids proceeds with development of uniform, continuous frontiers of reaction.However this point of view is doubtful / 1 /. E.g. we have proved the active role of the block (grain) boundaries in the main phases of cement, these boundaries being the areas of hydrate phases' nucleation / 2 /. It has brought to the supposition that the dissolution frontier of cement particles in water is discrete. It seems also probable that the dissolution proceeds through the channels, which serve both for the liquid phase movement and for the drainage of the incongruant solution products. These channels can be appeared along the block boundaries.In order to demonsrate it, we have offered the method of phase-contrast impregnation of the hardened cement paste with the solution of methyl metacrylahe and benzoyl peroxide. The viscosity of this solution is equal to that of water.


Author(s):  
William Krakow ◽  
Benjamin Siegel

Unwin has used a metallized non-conducting thread in the back focal plane of the objective lens that stops out a portion of the unscattered beam, takes on a localized positive charge and thus produces an additional phase shift to give a different transfer function of the lens. Under the particular conditions Unwin used, the phase contrast image was shifted to bright phase contrast for optimum focus.We have investigated the characteristics of this type of electrostatic phase plate, both analytically and experimentally, as functions of the magnitude of charge and defocus. Phase plates have been constructed by using Wollaston wire to mount 0.25μ diameter platinum wires across apertures ranging from 50 to 200μ diameter and vapor depositing SiO and gold on the mounted wires to give them the desired charging characteristics. The net charge was varied by adjusting only the bias on the Wehnelt shield of the gun, and hence the beam currents and effective size of the source.


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