Advanced Fixation for Transmission Electron Microscopy Unveils Special Extracellular Matrix Within the Renal Stem/Progenitor Cell Niche

2014 ◽  
pp. 21-37 ◽  
Author(s):  
Will W. Minuth ◽  
Lucia Denk
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Extracellular Matrix ◽  
Progenitor Cell ◽  
Transmission Electron ◽  
Cell Niche

Collagen Characteristics and Organization during the Progression of Cholesterol-Induced Atherosclerosis in Japanese Quail

2001 ◽  
Vol 226 (4) ◽  
pp. 328-333 ◽  
Author(s):  
Sandra G. Velleman ◽  
Richard J. McCormick ◽  
Daniel Ely ◽  
Bradley B. Jarrold ◽  
Ruthi A. Patterson ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Blood Pressure ◽  
Transmission Electron Microscopy ◽  
Extracellular Matrix ◽  
Systolic Blood Pressure ◽  
Japanese Quail ◽  
Collagen Fibril ◽  
Cholesterol Feeding ◽  
Transmission Electron ◽  
Progression Of Atherosclerosis

This study reports the concentration of collagen and its hydroxypyridinoline crosslinks, collagen fibril organization in the dorsal aortas, and systolic blood pressure during the progression of atherosclerosis in Japanese quail selected for cholesterol-induced atherosclerosis. The quail were placed on either a control or 0.5% cholesterol-added diet at approximately 16 weeks of age. The concentration of total collagen did not change in the control arteries during the course of the study, whereas at 5 and 10 weeks of cholesterol feeding, collagen levels decreased in the cholesterol-fed birds. Hydroxypyridinoline concentration increased during the duration of the study in the cholesterol-fed birds and by 15 and 20 weeks of cholesterol feeding, levels were significantly increased over those observed in the control arteries. Transmission electron microscopy showed changes in the organization of collagen fibrils. Increased systolic blood pressure was noted beginning at 10 weeks of cholesterol feeding, which is suggestive of other systemic changes induced by hypercholesterolemia. These results demonstrated remodeling of the collagen component of the dorsal aorta extracellular matrix during the progression of atherosclerosis and are suggestive of other systemic cardiovascular system changes.

Aspects ultrastructuraux de l'interaction uretère-mesenchyme, au cours de l'induction néphrogène, chez les amphibiens

Development ◽  
1977 ◽  
Vol 41 (1) ◽  
pp. 259-268
Author(s):  
Par J.-D. Gipouloux ◽  
M. Delbos
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Extracellular Matrix ◽  
Close Contact ◽  
Close Apposition ◽  
Collagen Fibres ◽  
Morphological Basis ◽  
Transmission Electron

Ultrastructural features of ureter-mesenchyme interaction, during nephrogenic induction in Amphibia Transmission electron microscopy was used to study the interface between the ureter and the mesonephric blastema. When the blastema is induced in a functional organ, numerous pseudopodial protrusions of the ureter and of the mesonephric cells lead to a close apposition between the two structures. The presence of squamous material, perhaps of a mucopolysaccharidic nature, with a network of collagen fibres is always found on this interface. These particular processes seem to play a role as a morphological basis for the phenomenon of induction. The importance of the extracellular matrix is discussed. Neither the occurrence of zones of close contact, reported by several authors, in other species, nor the fusion of membranes has ever been observed, in Amphibia.

scholarly journals A pre-embedding immunolabeling technique for basal lamina and extracellular matrix molecules.

1988 ◽  
Vol 36 (4) ◽  
pp. 453-458 ◽  
Author(s):  
M M Martins-Green ◽  
K T Tokuyasu
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Extracellular Matrix ◽  
Optical Microscopy ◽  
Basal Lamina ◽  
Serial Sections ◽  
Stages Of Development ◽  
Additional Advantage ◽  
Transmission Electron ◽  
Extracellular Matrix Molecules

We have developed a pre-embedding immunolabeling technique to identify basal lamina and extracellular matrix molecules in embryos at various stages of development. The technique works for both fluorescence optical microscopy (1-2.5-micron sections) and for transmission electron microscopy, and enables straigthforward correlation between the two. An additional advantage is the easy preparation of well-oriented serial sections, facilitating detailed studies of development.

The acellular stroma of the chick cornea inhibits melanogenesis of the neural-crest-derived cells that colonize it.

Development ◽  
1985 ◽  
Vol 86 (1) ◽  
pp. 143-154
Author(s):  
Steven Campbell ◽  
Jonathan B. L. Bard
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Extracellular Matrix ◽  
Neural Crest ◽  
Corneal Stroma ◽  
Cell Types ◽  
Brown Pigment ◽  
Transmission Electron ◽  
Wide Range

Neural crest (NC) cells from the periorbital mesenchyme (POM) invade the acellular stroma of the chick cornea at stage 27 of development (∼6 days). The invading cells become collagenproducing fibroblasts while the NC cells remaining in the POM differentiate into a wide range of cell types, the most easily recognizable of which is the pigment-producing melanocyte. In this paper, we report observations on the differentiation in vitro of cells within and migrating from explants of corneal stroma and compare their behaviour with that of cells within and migrating from explants of the POM. In ∼70% of cases, POM explants produced black, eumelanin pigmentation within 2–3 days in culture and gave rise to a mixed outgrowth of fibroblasts and melanoblasts that produced brown pigment. In no case, however, did a corneal explant produce black pigment (so demonstrating that any POM contamination was negligible). However, in 28% of cultures from stage-27 and -28 corneas, some of the cells in the outgrowth contained brown pigment indistinguishable from that produced by the POM control, although the majority of the cells in each case were fibroblasts. Two lines of investigation demonstrated that this pigment was melanin: first, transmission electron microscopy showed that the pigment organelles were incompletely melanized, granular melanosomes; second, tests designed to demonstrate the presence of lipofuscin, an alternative pigment, proved negative. Migrating cells from older corneas, in contrast, showed no evidence of even the first stages of melanogenesis. These results show, first, that some of the NC cells that invade the cornea are at least bipotent and hence representative of the POM population rather than being a unique subgroup and, second, that the acellular stroma of the cornea determines the state of differentiation of the NC cells that colonize it. The results thus provide an unequivocal demonstration that extracellular matrix can induce postmigratory NC cells to differentiate into fibroblasts.

scholarly journals Thrombospondin-2 deficiency in growing mice alters bone collagen ultrastructure and leads to a brittle bone phenotype

2015 ◽  
Vol 119 (8) ◽  
pp. 872-881 ◽  
Author(s):  
Eugene Manley ◽  
Joseph E. Perosky ◽  
Basma M. Khoury ◽  
Anita B. Reddy ◽  
Kenneth M. Kozloff ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mechanical Properties ◽  
Transmission Electron Microscopy ◽  
Extracellular Matrix ◽  
Cortical Bone ◽  
Type I Collagen ◽  
Type I ◽  
Wild Type ◽  
Transmission Electron ◽  
Series Of Experiments

Thrombospondin-2 (TSP2) is a matricellular protein component of the bone extracellular matrix. Long bones of adult TSP2-deficient mice have increased endosteal bone thickness due to expansion of the osteoblast progenitor cell pool, and these cells display deficits in osteoblastic potential. Here, we investigated the effects of TSP2 deficiency on whole bone geometric and mechanical properties in growing 6-wk-old male and female wild-type and TSP2-knockout (KO) mice. Microcomputed tomography and mechanical testing were conducted on femora and L2 vertebrae to assess morphology and whole bone mechanical properties. In a second series of experiments, femoral diaphyses were harvested from wild-type and TSP2-KO mice. Detergent-soluble type I collagen content was determined by Western blot of right femora. Total collagen content was determined by hydroxyproline analysis of left femora. In a third series of experiments, cortical bone was dissected from the anterior and posterior aspects of the femoral middiaphysis and imaged by transmission electron microscopy to visualize collagen fibrils. Microcomputed tomography revealed minimal structural effects of TSP2 deficiency. TSP2 deficiency imparted a brittle phenotype on cortical bone. Femoral tissue mineral density was not affected by TSP2 deficiency. Instead, transmission electron microscopy revealed less intensely stained collagen fibrils with altered morphology in the extracellular matrix assembled by osteoblasts on the anterior surface of TSP2-KO femora. Femoral diaphyseal bone displayed comparable amounts of total collagen, but the TSP2-KO bones had higher levels of detergent-extractable type I collagen. Together, our data suggest that TSP2 is required for optimal collagen fibrillogenesis in bone and thereby contributes to normal skeletal tissue quality.

Techniques for Transmission Electron Microscopy of Fiber-Matrix Interfaces in Aluminum Alloy-Boron Composites by Ion Erosio

1971 ◽  
Vol 29 ◽  
pp. 204-205
Author(s):  
G. G. Shaw
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Aluminum Alloy ◽  
Electron Beam ◽  
Pure Aluminum ◽  
Ion Milling ◽  
Transmission Electron ◽  
Fiber Matrix ◽  
Ion Erosion ◽  
Row Of Fibers

The morphology and composition of the fiber-matrix interface can best be studied by transmission electron microscopy and electron diffraction. For some composites satisfactory samples can be prepared by electropolishing. For others such as aluminum alloy-boron composites ion erosion is necessary.When one wishes to examine a specimen with the electron beam perpendicular to the fiber, preparation is as follows: A 1/8 in. disk is cut from the sample with a cylindrical tool by spark machining. Thin slices, 5 mils thick, containing one row of fibers, are then, spark-machined from the disk. After spark machining, the slice is carefully polished with diamond paste until the row of fibers is exposed on each side, as shown in Figure 1.In the case where examination is desired with the electron beam parallel to the fiber, preparation is as follows: Experimental composites are usually 50 mils or less in thickness so an auxiliary holder is necessary during ion milling and for easy transfer to the electron microscope. This holder is pure aluminum sheet, 3 mils thick.

Direct Observation of Heat Exchanger Deposits by Cross Sectioning

1967 ◽  
Vol 25 ◽  
pp. 364-365
Author(s):  
R. W. Anderson ◽  
D. L. Senecal
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Heat Exchanger ◽  
Direct Observation ◽  
Cross Sections ◽  
Preparation Method ◽  
Specimen Preparation ◽  
Flowing Water ◽  
Transmission Electron ◽  
Deposit Layer

A problem was presented to observe the packing densities of deposits of sub-micron corrosion product particles. The deposits were 5-100 mils thick and had formed on the inside surfaces of 3/8 inch diameter Zircaloy-2 heat exchanger tubes. The particles were iron oxides deposited from flowing water and consequently were only weakly bonded. Particular care was required during handling to preserve the original formations of the deposits. The specimen preparation method described below allowed direct observation of cross sections of the deposit layers by transmission electron microscopy.The specimens were short sections of the tubes (about 3 inches long) that were carefully cut from the systems. The insides of the tube sections were first coated with a thin layer of a fluid epoxy resin by dipping. This coating served to impregnate the deposit layer as well as to protect the layer if subsequent handling were required.

Phase Transformations in Ti-7w/oMo-3w/oMe Alloy

1974 ◽  
Vol 32 ◽  
pp. 514-515
Author(s):  
S. Fujishiro
Keyword(s):  
Electron Microscopy ◽  
Mechanical Properties ◽  
Transmission Electron Microscopy ◽  
Tensile Strength ◽  
Mechanical Processing ◽  
Ray Method ◽  
X Ray ◽  
Transmission Electron ◽  
Water Quench ◽  
Alpha Beta

The mechanical properties of three titanium alloys (Ti-7Mo-3Al, Ti-7Mo- 3Cu and Ti-7Mo-3Ta) were evaluated as function of: 1) Solutionizing in the beta field and aging, 2) Thermal Mechanical Processing in the beta field and aging, 3) Solutionizing in the alpha + beta field and aging. The samples were isothermally aged in the temperature range 300° to 700*C for 4 to 24 hours, followed by a water quench. Transmission electron microscopy and X-ray method were used to identify the phase formed. All three alloys solutionized at 1050°C (beta field) transformed to martensitic alpha (alpha prime) upon being water quenched. Despite this heavily strained alpha prime, which is characterized by microtwins the tensile strength of the as-quenched alloys is relatively low and the elongation is as high as 30%.

Two- or three-way observations of the identical cell elements within the same sections by LM, TEM and/or SEM

1978 ◽  
Vol 36 (2) ◽  
pp. 82-83 ◽  
Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Light Microscopy ◽  
Thin Sections ◽  
Transmission Electron ◽  
Identical Cell ◽  
Electron Microscopes ◽  
Scanning Electron

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).

Comparative Microstructures of Ion Milled and Electrochemically Thinned Composite Materials

1974 ◽  
Vol 32 ◽  
pp. 546-547
Author(s):  
R.R. Russell
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Composite Materials ◽  
Great Difficulty ◽  
Ion Milling ◽  
Transmission Electron ◽  
Ion Damage ◽  
Intermetallic Composite

Transmission electron microscopy of metallic/intermetallic composite materials is most challenging since the microscopist typically has great difficulty preparing specimens with uniform electron thin areas in adjacent phases. The application of ion milling for thinning foils from such materials has been quite effective. Although composite specimens prepared by ion milling have yielded much microstructural information, this technique has some inherent drawbacks such as the possible generation of ion damage near sample surfaces.

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