scholarly journals Three-dimensional interrelationship between osteocyte network and forming mineral during human bone remodeling

2020 ◽  
Author(s):  
Mahdi Ayoubi ◽  
Alexander F. van Tol ◽  
Richard Weinkamer ◽  
Paul Roschger ◽  
Peter C. Brugger ◽  
...  

AbstractDuring bone remodeling, osteoblasts are known to deposit unmineralized collagenous tissue (osteoid), which mineralizes after some time lag. Some of the osteoblasts differentiate into osteocytes, forming a cell network within the lacunocanalicular network (LCN) of bone. To get more insight into the potential role of osteocytes in the mineralization process of osteoid, sites of bone formation were three-dimensionally imaged in nine forming human osteons using focused ion beam-scanning electron microscopy (FIB-SEM). In agreement with previous observations, the mineral concentration was found to gradually increase from the central Haversian canal towards preexisting mineralized bone. Most interestingly, a similar feature was discovered on a length scale more than 100-times smaller, whereby mineral concentration increased from the LCN, leaving around the canaliculi a zone virtually free of mineral, the size of which decreases with progressing mineralization. This suggests that the LCN controls mineral formation but not just by diffusion of mineralization precursors, which would lead to a continuous decrease of mineral concentration from the LCN. Our observation is, however, compatible with the codiffusion and reaction of precursors and inhibitors from the LCN into the bone matrix.

Author(s):  
T. Yaguchi ◽  
M. Konno ◽  
T. Kamino ◽  
M. Ogasawara ◽  
K. Kaji ◽  
...  

Abstract A technique for preparation of a pillar shaped sample and its multi-directional observation of the sample using a focused ion beam (FIB) / scanning transmission electron microscopy (STEM) system has been developed. The system employs an FIB/STEM compatible sample rotation holder with a specially designed rotation mechanism, which allows the sample to be rotated 360 degrees [1-3]. This technique was used for the three dimensional (3D) elemental mapping of a contact plug of a Si device in 90 nm technology. A specimen containing a contact plug was shaped to a pillar sample with a cross section of 200 nm x 200 nm and a 5 um length. Elemental analysis was performed with a 200 kV HD-2300 STEM equipped with the EDAX genesis Energy dispersive X-ray spectroscopy (EDX) system. Spectrum imaging combined with multivariate statistical analysis (MSA) [4, 5] was used to enhance the weak X-ray signals of the doped area, which contain a low concentration of As-K. The distributions of elements, especially the dopant As, were successfully enhanced by MSA. The elemental maps were .. reconstructed from the maps.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiyu Sun ◽  
Wei Wu ◽  
Limei Tian ◽  
Wei Li ◽  
Fang Zhang ◽  
...  

AbstractNot only does the Dynastes tityus beetle display a reversible color change controlled by differences in humidity, but also, the elytron scale can change color from yellow-green to deep-brown in specified shapes. The results obtained by focused ion beam-scanning electron microscopy (FIB-SEM), show that the epicuticle (EPI) is a permeable layer, and the exocuticle (EXO) is a three-dimensional photonic crystal. To investigate the mechanism of the reversible color change, experiments were conducted to determine the water contact angle, surface chemical composition, and optical reflectance, and the reflective spectrum was simulated. The water on the surface began to permeate into the elytron via the surface elemental composition and channels in the EPI. A structural unit (SU) in the EXO allows local color changes in varied shapes. The reflectance of both yellow-green and deep-brown elytra increases as the incidence angle increases from 0° to 60°. The microstructure and changes in the refractive index are the main factors that influence the process of reversible color change. According to the simulation, the lower reflectance causing the color change to deep-brown results from water infiltration, which increases light absorption. Meanwhile, the waxy layer has no effect on the reflection of light. This study lays the foundation to manufacture engineered photonic materials that undergo controllable changes in iridescent color.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexey A. Polilov ◽  
Anastasia A. Makarova ◽  
Song Pang ◽  
C. Shan Xu ◽  
Harald Hess

AbstractModern morphological and structural studies are coming to a new level by incorporating the latest methods of three-dimensional electron microscopy (3D-EM). One of the key problems for the wide usage of these methods is posed by difficulties with sample preparation, since the methods work poorly with heterogeneous (consisting of tissues different in structure and in chemical composition) samples and require expensive equipment and usually much time. We have developed a simple protocol allows preparing heterogeneous biological samples suitable for 3D-EM in a laboratory that has a standard supply of equipment and reagents for electron microscopy. This protocol, combined with focused ion-beam scanning electron microscopy, makes it possible to study 3D ultrastructure of complex biological samples, e.g., whole insect heads, over their entire volume at the cellular and subcellular levels. The protocol provides new opportunities for many areas of study, including connectomics.


2007 ◽  
Vol 15 (2) ◽  
pp. 26-31 ◽  
Author(s):  
Ben Lich

DualBeam instruments that combine the imaging capability of scanning electron microscopy (SEM) with the cutting and deposition capability of a focused ion beam (FIB) provide biologists with a powerful tool for investigating three-dimensional structure with nanoscale (1 nm-100 nm) resolution. Ever since Van Leeuwenhoek used the first microscope to describe bacteria more than 300 years ago, microscopy has played a central role in scientists' efforts to understand biological systems. Light microscopy is generally limited to a useful resolution of about a micrometer. More recently the use of confocal and electron microscopy has enabled investigations at higher resolution. Used with fluorescent markers, confocal microscopy can detect and localize molecular scale features, but its imaging resolution is still limited. SEM is capable of nanometer resolution, but is limited to the near surface region of the sample.


Author(s):  
W. N. P. Hung ◽  
M. M. Agnihotri ◽  
M. Y. Ali ◽  
S. Yuan

Traditional micromanufacturing has been developed for semiconductor industry. Selected micro electrical mechanical systems (MEMS) have been successfully developed and implemented in industry. Since current MEMS are designed for manufacture using microelectronics processes, they are limited to two-dimensional profiles and semiconductor based materials. Such shape and material constraints would exclude many applications that require biocompatibility, dynamic stress, and high ductility. New technologies are sought to fabricate three dimensional microcomponents using robust materials for demanding applications. To be cost effective, such microdevices must be economically mass producible. Molding is one of the promising replication techniques to mass produce components from polymers and polymer-based composites. This paper presents the development of a micromolding process to produce thermoplastic microcomponents. Mold design required precision fitting and was integrated with a vacuum pump to minimize air trap in mold cavities. Nickel and aluminum mold inserts were used for the study; their cavities were fabricated by combinations of available micromachining processes like laser micromachining, micromilling, micro electrical discharge machining, and focused ion beam sputtering. High and low density polyethylene, polystyrene polymers were used for this study. The effects of polymer molecular structures, molding temperature, time, and pressure on molding results were studied. Simulation of stress in the microcomponents, plastic flow in microchannels, and mold defects was performed and compare with experimental data. The research results showed that a microcomponent can be fabricated to the minimum size of 10 ± 1μm (0.0004 inch) with surface roughness <10 nm Rt. Molding of micro-size geartrains and orthopedic meso-size fasteners was completed to illustrate the capability of this process.


Contact ◽  
2018 ◽  
Vol 1 ◽  
pp. 251525641880142
Author(s):  
Manon Rosselin ◽  
Paula Nunes-Hasler ◽  
Nicolas Demaurex

Mitochondria undergo spontaneous transient elevations in matrix pH associated with drops in mitochondrial membrane potential. These mitopHlashes require a functional respiratory chain and the profusion protein optic atrophy 1, but their mechanistic basis is unclear. To gain insight on the origin of these dynamic events, we resolved the ultrastructure of flashing mitochondria by correlative light and electron microscopy. HeLa cells expressing the matrix-targeted pH probe mitoSypHer were screened for mitopHlashes and fixed immediately after the occurrence of a flashing event. The cells were then processed for imaging by serial block face scanning electron microscopy using a focused ion beam to generate ∼1,200 slices of 10 nm thickness from a 28 µm × 15 µm cellular volume. Correlation of live/fixed fluorescence and electron microscopy images allowed the unambiguous identification of flashing and nonflashing mitochondria. Three-dimensional reconstruction and surface mapping revealed that each tomogram contained two flashing mitochondria of unequal sizes, one being much larger than the average mitochondrial volume. Flashing mitochondria were 10-fold larger than silent mitochondria but with a surface to volume ratio and a cristae volume similar to nonflashing mitochondria. Flashing mitochondria were connected by tubular structures, formed more membrane contact sites, and a constriction was observed at a junction between a flashing mitochondrion and a nonflashing mitochondrion. These data indicate that flashing mitochondria are structurally preserved and bioenergetically competent but form numerous membrane contact sites and are connected by tubular structures, consistent with our earlier suggestion that mitopHlashes might be triggered by the opening of fusion pores between contiguous mitochondria.


2017 ◽  
Vol 23 (5) ◽  
pp. 916-925
Author(s):  
Pritesh Parikh ◽  
Corey Senowitz ◽  
Don Lyons ◽  
Isabelle Martin ◽  
Ty J. Prosa ◽  
...  

AbstractThe semiconductor industry has seen tremendous progress over the last few decades with continuous reduction in transistor size to improve device performance. Miniaturization of devices has led to changes in the dopants and dielectric layers incorporated. As the gradual shift from two-dimensional metal-oxide semiconductor field-effect transistor to three-dimensional (3D) field-effect transistors (finFETs) occurred, it has become imperative to understand compositional variability with nanoscale spatial resolution. Compositional changes can affect device performance primarily through fluctuations in threshold voltage and channel current density. Traditional techniques such as scanning electron microscope and focused ion beam no longer provide the required resolution to probe the physical structure and chemical composition of individual fins. Hence advanced multimodal characterization approaches are required to better understand electronic devices. Herein, we report the study of 14 nm commercial finFETs using atom probe tomography (APT) and scanning transmission electron microscopy–energy-dispersive X-ray spectroscopy (STEM-EDS). Complimentary compositional maps were obtained using both techniques with analysis of the gate dielectrics and silicon fin. APT additionally provided 3D information and allowed analysis of the distribution of low atomic number dopant elements (e.g., boron), which are elusive when using STEM-EDS.


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