The morphology and composition of cholesterol,protein, and bilirubin deposits in dried human bile: Cathodoluminescence and backscattered electron imaging

This work is the first to deal with the application of color cathodoluminescence scanning electron microscopy (CCL SEM) and a novel version of combined imaging with backscattered electrons (CCL+BSE SEM) for the study of the composition of bile and its precipitation mechanisms. The present study demonstrates cholesterol, protein, and bilirubin distribution in deposits of normal and abnormal humanbile after solution evaporation to full dryness. Qualitative CCL SEM analysis showed that dried bile remnants include different proportions of the above components. Three types of deposits were observed: Arborescent crystals, typical cholesterol crystals, and amorphous bilirubin particles. The selection of crystalline or amorphous precipitate phases is determined by the dehydration/concentration process. The findings may explain key features in lithogenesis.     

Huge clusters of embryonic stem cells in human embryos: A morphologic study

Background: Nothing is known about huge clusters (HC) of embryonic stem cells (ESC) in human fetal organs (HFO). Aim: To know the status of HC‐ESC in HFO. Methods: Morphology and immunohistochemistry (IHC) in 32 HFO of 7–40 gestational weeks (GW). Results: HC‐ESC were seen in many HFO including central nervous system, spinal cords, spine, soft tissue, bone, skin, thyroid, lung, liver, pancreas, gall bladder, extrahepatic bile duct, adrenal, kidney, bladder, foregut, midgut, hindgut, female and male genital organs, and neurons. HC‐ESC's were composed of two populations depending on constituting cells. One were large cells with ample acidophilic cytoplasms with vesicular nuclei and nucleoli. The other were small cells with scant cytoplasm with hyperchromatic nuclei without nucleoli, resembling lymphocytes. The HC‐ESC were frequently showed neuronal differentiation. HC‐ESC were positive for NCAM, synaptophysin, NSE, chromogranin, PDGFRA, AFP, ErbB2, bcl‐2, KIT, MET. They were negative for CD45, CD3, CD20, EMA, CEA, CA19‐9, cytokeratin (CK) 7, CK8, CK18, CK19, MUC1, MUC2, MUC5AC, and MUC6. The mean Ki‐67 labeling index (LI) was 13% ± 7%. HC‐ESC showed a little glycogen but lacked mucins. These HC‐ESC were seen in 7–25 GW, and they were rarely seen in 26–40 GW. Conclusions: The morphology, IHC, and ontogeny of HC‐ESC were described. Microsc. Res. Tech. 77:825–831, 2014. © 2014 Wiley Periodicals, Inc.     

Development of spiral ganglion cells in mammalian cochlea

The development of the spiral ganglion in the cat, the rat, and the mouse was studied by electron microscopy, from fetal stages in the cat and from birth in the rodent. In the earliest stages, a single population of ganglion cells is present. Immature spiral ganglion neurons possess small perisomatic processes that seem to disappear with development, before the myelination ganglion cells are surrounded by one or two layers of Schwann cell processes. With maturation, the Schwann process increases in number around the perikaryon and its processes, which leads to the onset of myelination. The onset of myelination of the cell body processes is asynchronous. The perikaryon may be delayed in myelination by several days. Moreover, ganglion neurons from a given region of the cochlea do not myelinate simultaneously. The differentiation of two types of fibers in the intraganglionic spiral bundle and the first appearance of TII neurons occurs around birth in the cat and a few days after birth for the rat and the mouse. The distinction of TII cells is possible due to characteristic accumulation of neurofilamentous structures in the cytoplasm.     

Tracing axons in the human brain: A method utilizing light and TEM techniques

The neuroanatomy of experimental animals has been investigated through a variety of histological and physiological techniques. Recently, more sophisticated methods have permitted a better understanding of the complex connections of animal brains. In contrast, the methods available for the direct examination of the human brain have remained relatively primitive. We have developed a staining method (paraphenylene-diamine method: PPD) which bridges the techniques of light and electron microscopy. The method permits the tracing of degenerated fibers in the human brain even after very long survival periods. With this method we have documented several visual pathways not previously described in man. We have also demonstrated, in several species, that products of axonal degeneration remain far longer than previously supposed. The PPD technique is relatively simple, reliable, provides high resolution, works in human brains, and can be used in conjunction with TEM.     

High-resolution X-ray tomography of the human inner ear: synchrotron radiation-based study of nerve fibre bundles, membranes and ganglion cells

The combination of osmium tetroxide staining and high-resolution tomographic imaging using monochromatic X rays allows visualizing cellular structures of the human inner ear, that is, the organ of Corti, the stria vascularis and further soft tissues of the membranous labyrinth, in three-dimensional space with isotropic micrometre resolution. This approach permits to follow the course of nerve fibre bundles in a major part of the specimen and reveals the detailed three-dimensional arrangement of individual ganglion cells with distinct nuclei by means of X-ray tomography for the first time. The non-destructive neuron cell counting in a selected volume of 125 μm × 800 μm × 600 μm = 0.06 mm³ gives rise to the estimate that 2000 ganglion cells are present along 1 mm organ of Corti.     

Multifractal characterization of morphology of human red blood cells membrane skeleton

The purpose of this paper is to show applicability of multifractal analysis in investigations of the morphological changes of ultra‐structures of red blood cells (RBCs) membrane skeleton measured using atomic force microscopy (AFM). Human RBCs obtained from healthy and hypertensive donors as well as healthy erythrocytes irradiated with neutrons (45 μGy) were studied. The membrane skeleton of the cells was imaged using AFM in a contact mode. Morphological characterization of the three‐dimensional RBC surfaces was realized by a multifractal method. The nanometre scale study of human RBCs surface morphology revealed a multifractal geometry. The generalized dimensions D_q and the singularity spectrum f(α) provided quantitative values that characterize the local scale properties of their membrane skeleton organization. Surface characterization was made using areal ISO 25178‐2: 2012 topography parameters in combination with AFM topography measurement. The surface structure of human RBCs is complex with hierarchical substructures resulting from the organization of the erythrocyte membrane skeleton. The analysed AFM images confirm a multifractal nature of the surface that could be useful in histology to quantify human RBC architectural changes associated with different disease states. In case of very precise measurements when the red cell surface is not wrinkled even very fine differences can be uncovered as was shown for the erythrocytes treated with a very low dose of ionizing radiation.     

Three-dimensional reconstruction of the human renal glomerulus

The capillary bed of the human renal glomerulus is one of the more complex capillary structures in the human body. This paper illustrates three-dimensional reconstruction of the capillary bed from serial sections. It shows that, although traditional methods of three-dimensional rendering by computer fail to handle the complexities of the capillary structure, new methods based on filtering using three-dimensional mathematical morphology are capable of revealing previously unseen details. This is done at the expense of eliminating fine structure (small capillaries). An error analysis allows the degree to which fine details are lost to be estimated.     

Taxonomic importance of spore morphology in Thelypteridaceae from Northern Pakistan

Spore morphology of Thelypteridaceae species growing in Malakand Division, Northern Pakistan, was studied using both light microscopy and scanning electron microscopy. The taxa are Christella dentata and Glaphyropteridopsis erubescens in the subfamily Thelypteridoideae, and Phegopteris connectilis, Pseudophegopteris pyrrhorhachis, and Pseudophegopteris levingei in the subfamily Phegopteridoideae. The studied species exhibit differences in spore size, exospore thickness, color, and ornamentation. Spores of the studied species are monolete and medium‐sized, and shape is ellipsoidal in both polar and equatorial views. The average measurement of the polar diameter ranges from 27 μm to 31 μm, whereas in the equatorial direction it varied from 20 μm to 40 μm. The exospore thickness ranges from 1.2 μm to 2.4 μm. Reticulate, laevigate with microgranules, cristate, and coarsely echinate surface ornamentation are observed among the species. Multivariate analysis including unweighted pair group method with arithmetic mean and principal component analysis was used for the grouping and discrimination of species and genera.     

On predicting chip morphology and phase transformation in hard machining

A finite element model is developed to predict the chip formation and phase transformation in orthogonal machining of hardened AISI 52100 steel (62HRC) using Polycristalline Cubic Boron Nitride (PCBN) tools. The model mainly includes a chip separation criterion based on critical equivalent plastic strain; a Coulomb’s law for the friction at the tool/chip interface; a material constitutive relation of velocity-modified temperature; a thermal analysis incorporating the heat dissipated from inelastic deformation energy and friction; and an annealing effect model, in which the work hardening effect may be lost or re-accumulate depending on material temperature. This fully coupled thermal-mechanical finite element analysis accurately simulates the formation of segmental chips and predicts the phase transformation on the chips, as verified by experiment. It is found that high temperatures around the secondary shear zone causes fast re-austenitization and martensite transformation, while other parts of the chips retain the original tempered martensitic structure.     

Multidimensional long-term time-lapse microscopy of in vitro peripheral nerve regeneration

In order to test the effectiveness of a new advanced time-lapse microscopy imaging and image processing and analysis system, and to do quantitative and qualitative temporal analyses of in vitro peripheral nerve regeneration, long-term time-lapse imaging of cultures of mouse dorsal root ganglia (DRGs) was performed. DRGs were placed in a Petri dish, covered with collagen gel, their attached peripheral nerves were cut in the middle, creating a gap, and the dish was filled with culture medium. Six preparations were kept on the time-lapse imaging system, which provides a suitable incubation environment and enables to capture images from multiple coordinates at x,y,z axes at desired time intervals for 13 days. In general, the time-lapse imaging system proved quite stable and efficient, although some improvements are certainly required. Two main components of peripheral nerve regeneration, outgrowth of axons and activities of resident cells, were examined. Axons started to grow during the first hour of incubation with a 16.5 microm/h rate and showed the slowest rates (0.7 microm/h) on days 8 and 9, after which they resumed higher speeds again. The first cell came out of the proximal end of the cut nerve on the second day and it was a Schwann cell (SC), which was the prominent cell type in the preparations throughout the experiment. SCs were higher in number (83.15% of all cells) but slower in migration (3.4 vs. 7.3 microm/h, P < 0.001) than other cells. Other observed characteristics of axonal outgrowth and cellular activity and interactions between axons and the cells are discussed.     

Auditory hair cells in human fetuses: Synaptogenesis and ciliogenesis

This paper brings together the most recent findings concerning the development of human fetus cochlear hair cells, as observed using transmission and scanning electron microscopy (TEM and SEM). Specific attention is drawn to the formation of synapses and the growth of stereocilia. In both types of hair cells, synaptogenesis begins before ciliogenesis (week 10 of gestation versus week 12). In the inner hair cell (IHC), an adultlike stage is seen around week 15 for synapses, but not before week 22 for stereocilia. In the outer hair cell (OHC), both synapses and stereocilia are not yet fully mature at week 22. Classic gradients of maturation are found: a base-to-apex gradient, and an IHC-to-OHC gradient. By comparing these results with the anatomical and functional data on cochlear development in laboratory mammals, the onset of cochlear function in the human fetus can be estimated to occur around week 18. The completion of cochlear maturation based upon the same anatomical criteria should occur about 10 weeks later.     

Morphological observations of small granule-containing (Chromaffin) cells in the celiac ganglion of the guinea pig,with emphasis on cell contacts

Utilizing electron microscopic observation, several contacts between small, granule-containing cells (SGC) and postganglionic neurons (PGN) in the celiac ganglion of the guinea pig have been observed. A SGC in very close association with a PGN was seen to receive a distinct synaptic contact that contained many vesicles with dense cores. This contact was morphologically unlike cholinergic synapses previously reported on chromaffin cells. Because the SGC and PGN were clearly separated by a thin rim of satellite cell cytoplasm mutual to both cells, it is not known how or if the SGC would possibly exert a synaptic or paracrine effect on the PGN. Also, intraganglion SGC existed as large well-vascularized islands within the celiac ganglion. These intraganlion clusters sometimes contained more than 50 cells and perhaps could be considered to function as localized neuroendocrine components within the ganglion by secreting granule products into the nearby blood vessels for local or distant effects, although this certainly is not known. This work reports a unique synaptic ending upon a single-occurring SGC, which, in turn, closely approximates a ganglion neuron in a soma-somatic relationship. In addition, a very close association (but no actual contact) was observed between granule-containing processes, presumably emanating from the intraganglion clusters, and PGN. Whatever the function of ganglionic SGC may be, the exact relationship between SGC and PGN presumably would be of great interest and potential importance. © 1994 Wiley-Liss, Inc.     

BGA焊点空洞缺陷的数字形态学检测

空洞缺陷是BGA焊接缺陷中比较常见的一种, 主要由回流焊时产生的气体没有及时排出而导致. X射线无损检测技术可以使空洞缺陷在焊球图像上显示为白色区域供技术人员查看, 但在噪声、不均匀照射、存在与缺陷类似目标的干扰下如何准确地自动提取缺陷一直是个难题. 提出使用Otsu算法分割焊球, 使用数学形态学中的开闭运算、 顶帽变换提取空洞缺陷的技术方案, 实现每个焊点空洞缺陷的准确提取和自动分析. 实验结果表明, 提出的技术方案鲁棒性强, 可应用于BGA焊点的空洞缺陷检测.     

Stem cells in the development and differentiation of the human adrenal glands

There are no studies on stem cells (SCs) and development and differentiation (DD) of the human adrenal glands. The SCs in DD of the adrenal glands were herein investigated histochemically and immunohistochemically in 18 human embryonic adrenal glands at gestational week (GW) 7–40. At 7 GW, the adrenal glands were present, and at 7 GW, numerous embryonic SCs (ESCs) are seen to create the adrenal cortex. The ESCs were composed exclusively of small cells with hyperchromatic nuclei without nucleoli. The ESCs were positive for neural cell adhesion molecule, KIT, neuron‐specific enolase, platelet‐derived growth factor receptor‐α, synaptophysin, and MET. They were negative for other SC antigens, including chromogranin, ErbB2, and bcl‐2. They were also negative for lineage antigens, including cytokeratin (CK)7, CK8, CK18, and CK19, carcinoembryonic antigen, carbohydrate antigen 19‐9, epithelial membrane antigen, HepPar1, mucin core apoprotein (MUC)1, MUC2, MUC5AC, and MUC6, and cluster differentiation (CD)3, CD45, CD20, CD34, and CD31. The Ki‐67 labeling index (LI) was high (Ki‐67 LI = around 20%). α‐Fetoprotein was positive in the ESCs and adrenal cells. The ESC was first seen in the periphery of the adrenal cortex at 7–10 GW. The ESC migrates into the inner part of the adrenal cortex. Huge islands of ESC were present near the adrenal, and they appeared to provide the ESC of the adrenal. At 16 GW, adrenal medulla appeared, and the adrenal ESCs were present in the periphery or the cortex, in the cortical parenchyma, corticomedullary junctions, and in the medulla. The adrenal essential architecture was established around 20 GW; however, there were still ESCs. At term, there are a few ESCs. These data suggest that the adrenal glands were created by ESCs. Microsc. Res. Tech., 78:59–64, 2015. © 2014 Wiley Periodicals, Inc.     

Surface morphology in engineering applications: Influence of roughness on sliding and wear in dry fretting

Influence of initial surface roughness on friction and wear processes under fretting conditions was investigated experimentally. Rough surfaces (Ra=0.15-2.52 μm) were prepared on two materials: carbon alloy (AISI 1034) and titanium alloy (Ti-6Al-4V). Strong influence of initial surface roughness on friction and wear processes is reported for both tested materials. Lower coefficient of friction and increase in wear rate was observed for rough surfaces. Wear activation energy is increasing for smoother surfaces. Lower initial roughness of surface subjected to gross slip fretting can delay activation of wear process and reduce wear rate; however, it can slightly increase the coefficient of friction.     

Cluster identification in AA5754 aluminium sheets using mathematical morphology analysis

Quantitative image analysis of particle distribution in the microstructure of continuous cast (CC) and direct chill cast (DC) AA5754 aluminium alloy sheets have been conducted. This information can be used as an input for modelling mechanical deformation and instability in these materials. The quantitative analysis reveals that there are significant differences in the microstructure of the two materials even though the total content of second‐phase particles is statistically similar. Qualitative observation shows the second‐phase particles to be arranged in the form of streaks parallel to the rolling direction in the CC sheets and in a uniform random manner in the DC sheets. The main difference in the geometric microstructure of the CC and DC material is the spatial arrangement of the second‐phase particles. A new mathematical technique called proximity analysis is developed to identify clusters and group of particles belonging to a cluster. Quantification through proximity analysis reveals that the particle clusters in CC sheet are in the form of long clusters (streaks) parallel to the rolling direction and are significantly longer than those in DC sheets (with the largest cluster in CC being four times larger than DC), and also have anisotropic angular orientation parallel to the rolling direction. The lower value of fracture strain observed in the CC sheets compared to DC sheets is attributed to a combination of large sizes of clusters and their preferential alignment along the rolling direction in the CC microstructure.     

Taxonomic importance of pollen morphology in Veratrum L. (Melanthiaceae) using microscopic techniques

Previous studies reported that intrageneric relationships of genus Veratrum of family Melanthiaceae are controversial and hard to delimit. Therefore, we observed the pollen morphological features of six species in the genus Veratrum in detail using both light microscopy and scanning electron microscopy and investigated their significance for Veratrum taxonomy. Among them, five were studied for the first time. The results demonstrated that pollen grains of Veratrum are medium in size with P/E being 0.31–0.60. Three types of shape in polar view have been observed elliptic, long‐elliptic, or wide‐elliptic with blunt or rounded at both ends. Two types of width of colpus reported narrow or wide, two types of depth of colpus reported deep or flat, and two types of length of colpus reported extend almost or do not extend to the ends, whereas two types of colpus membranes reported absent or obvious. One type of surface ornamentation was noted as reticulate. These results support species Veratrum album and Veratrum lobelianum as well as Veratrum grandiflorum and Veratrum oxysepalum as two independent species, respectively, rather than classifying Veratrum into two sections. Overall, we demonstrated that the ratio of polar axis length to equatorial axial length, pollen characteristics at the polar view, the colpus morphology, and the surface ornamentation of pollen grains of genus Veratrum have important systematic significance in identification and delimitation of species.     

A simple procedure for demonstrating the overall morphology of fibroblasts in routine histological preparations of adult tissues,using silver impregnation

A silver impregnation procedure for the total visualization of fibroblasts, with all their processes, is described. The method is applicable to routine formalin-fixed paraffin sections, or to formalin-fixed or native cryostat sections. The results are illustrated with examples from loose and dense connective tissue. Fibroblasts are visualized as cells with long, branching dendritic processes. The fibroblasts maintain contact with each other via these processes, thus forming a reticulum of cells, even in adult connective tissue.     

基于数学形态学的新方法在脑组织分割中的应用

针对人体脑部切片图像特点,提出了一种基于数学形态学的脑组织自动分割算法.该算法首先通过形态重构获得粗糙的脑组织区域,然后运用腐蚀和膨胀运算进行边界定位分割出了脑组织,最后对连续断层图像的分割结果进行了三维重建.结果表明该算法分割准确且自动化程度高,适合于大量序列切片图像的快速自动分割.     

磷酸二氢钾晶体飞切过程中温度场的分布及其对切屑形貌的影响

研究了磷酸二氢钾(KDP)晶体飞切加工过程中温度场的分布,探索了切削温度对KDP晶体切削过程的影响。首先,采用热力耦合有限元分析对KDP晶体切削过程进行了仿真,获得了不同切削深度下材料内部温度场的分布。分别使用飞切机床和纳米压痕仪在不同速度下切削KDP晶体,发现不同切削速度下形成的切屑的微观形貌存在显著差异,分析指出这可能是由于在不同切削速度下切削区域温度差异导致的。最后,对低速加工过程中获得的切屑进行加热试验,并观测了不同温升条件下切屑微观形貌的变化。飞切加工仿真实验显示:当切深为200nm时,切削区域的温度达到110℃;而实际实验结果表明:当温度超过100℃时,切屑的微观形貌会发生明显变化。综合仿真及实验结果可知:在KDP晶体飞切加工过程中切削区域的温度将超过100℃,因此在对KDP晶体切削机理进行研究时,必须考虑温度对材料力学性能及其去除过程的影响。