Microscopic radiography: A combined technique for improved analytical microscopic analysis and interpretration

1996 ◽  
Vol 54 ◽  
pp. 248-249
Author(s):  
A. Angel ◽  
R.C. Moretz
Keyword(s):  
Microscopic Analysis ◽  
Device Structure ◽  
Electron Microscopic ◽  
Drug Products ◽  
Microscopic Evaluation ◽  
Final Interpretation ◽  
And Function ◽  
Non Destructive ◽  
Combined Technique ◽  
Selection Of

Microscopic analysis of pharmaceutical devices and products relies primarily on destructive sample preparative methods, including sectioning, sawing, grinding and embedding. Reconstruction of images obtained from the prepared samples is often necessary to interpret the results. The preparative methods can introduce artifacts or distortion, which can affect the analysis, and the inability to visualize the intact object can also affect the interpretation. Radiography has been used to assist in the selection of preparative methods for microscopy, determine optimal orientation during preparation and analysis and to aid in the visualization and integration of the microscopic results in the final interpretation. The application of x-ray radiography to the examination of intact devices or manufactured drug products in conjunction with standard light and scanning electron microscopic evaluation presents a novel non-destructive technique to assess device structure and function as well as locate potential inclusions in manufactured drug products.

Electron microscopic analysis of the microtubular structure of HEp-2 cells

1990 ◽  
Vol 48 (3) ◽  
pp. 470-471
Author(s):  
Miklós S.Z. Kellermayer ◽  
Tamás Henics ◽  
György Szücs ◽  
Gerald H. Pollack
Keyword(s):  
Microscopic Analysis ◽  
Unknown Origin ◽  
Tubular Structure ◽  
Minimal Essential Medium ◽  
Electron Microscopic ◽  
Brij 58 ◽  
Cacodylate Buffer ◽  
Microscopic Studies ◽  
And Function ◽  
Hank’S Solution

The HEp-2 cell line was first established by Moore, et al. and today it serves as a widely used subject for a large variety of microbiological and cell-biological experiments. A tubular structure of unknown origin and function has been described first in virally infected, and later in uninfected HEp-2 cells. However, this tubular structure has not been further analyzed. We performed transmission (TEM) and whole cell mount electron microscopic studies of monolayer HEp-2 cells to morphologically describe the structure and relate it to other organelles of the cell.For embedding, HEp-2 cells were grown on glass coverslips in Eagle's minimal essential medium. 24 hours after passage, three types of procedures were carried out: (1) The cells were fixed with 2.5% glutaraldehyde in 0.14 M Na-cacodylate buffer. (2) The cells were treated for 10 min. with Hank's solution containing 0.2% Brij-58 and 2.5% glutaraldehyde. Fixation was completed with 2.5% glutaraldehyde in 0.14 M Na-cacodylate. (3) Cells were treated with 0.2% Brij-58 in Hank’s solution for 10 min. Subsequently, they were fixed with 2.5% glutaraldehyde in 0.14 M Na-cacodylate.

Desmosomal localization of beta-catenin in the skin of plakoglobin null-mutant mice

Development ◽  
1999 ◽  
Vol 126 (2) ◽  
pp. 371-381 ◽  
Author(s):  
C. Bierkamp ◽  
H. Schwarz ◽  
O. Huber ◽  
R. Kemler
Keyword(s):  
Heart Defects ◽  
Microscopic Analysis ◽  
Beta Catenin ◽  
Null Mutant ◽  
Electron Microscopic ◽  
Developmental Defects ◽  
Keratin Filaments ◽  
Embryonic Skin ◽  
Null Mutant Mice ◽  
And Function

Plakoglobin, a protein belonging to the Armadillo-repeat gene family, is the only component that adherens junctions and desmosomes have in common. Plakoglobin null-mutant mouse embryos die because of severe heart defects and may exhibit an additional skin phenotype, depending on the genetic background. Lack of plakoglobin affects the number and structure of desmosomes, resulting in visible defects when cells are subjected to increasing mechanical stress, e.g. when embryonic blood starts circulating or during skin differentiation. By analysing plakoglobin-negative embryonic skin differentiation in more detail, we show here that, in the absence of plakoglobin, its closest homologue, beta-catenin, becomes localized to desmosomes and associated with desmoglein. This substitution may account for the relatively late appearance of the developmental defects seen in plakoglobin null-mutant embryos. beta-catenin cannot, however, fully compensate a lack of plakoglobin. In the absence of plakoglobin, there was reduced cell-cell adhesion, resulting in large intercellular spaces between keratinocytes, subcorneal acantholysis and necrosis in the granular layer of the skin. Electron microscopic analysis documented a reduced number of desmosomes, and those present lacked the inner dense plaque and had fewer keratin filaments anchored. Our analysis underlines the central role of plakoglobin for desmosomal assembly and function during embryogenesis.

scholarly journals Influence of dark deprivation on the morphology of lamellarian bodies and other cell components of epidermis keratinocytes

2021 ◽  
Vol 18 (1) ◽  
pp. 80-88
Author(s):  
I. S. Sobolevskaya ◽  
O. D. Myadelets ◽  
O. B. Ostrovskaya
Keyword(s):  
Production Capacity ◽  
Microscopic Analysis ◽  
Epidermal Keratinocytes ◽  
Lamellar Bodies ◽  
Electron Microscopic ◽  
Microscopic Evaluation ◽  
The Status ◽  
Cell Components ◽  
Morphometric Assessment ◽  
Morphometric Evaluation

The aim of this study is to study the effect of dark deprivation on the ultrastructure of epidermal keratinocytes. Electron microscopic evaluation of the status of keratinocytes of the epidermis of the skin of white outbred rats with dark deprivation. For morphometric evaluation of the production capacity of counting the number of granules per 100 μm2. Using the application program ImageScopeM determines the average and equivalent indicators of lamellar bodies (μm), as well as their perimeter (μm); the average area of one section of the lalellar body (μm2); roundness coefficient and average relative electronic number of lamellar bodies.Electron microscopic analysis of keratinocytes revealed significant changes in their ultrastructure, which are due to the influence of desynchronosis. Maximum transformations were observed in Odland granules (lamellar bodies). Thus, the morphometric assessment revealed changes in the number (increase), their size (decrease), as well as in the distribution of lamellar plaques in them.With desynchronosis, significant changes in the ultrastructure of keratinocytes are observed. This indicates significant systemic disorders of the epidermis as a whole. In this transformation, there are cells of a special prickly and granular layer, in particular, lamellar bodies.

scholarly journals Immunochemical characterization of three components of the hemidesmosome and their expression in cultured epithelial cells.

1989 ◽  
Vol 109 (6) ◽  
pp. 3377-3390 ◽  
Author(s):  
D H Klatte ◽  
M A Kurpakus ◽  
K A Grelling ◽  
J C Jones
Keyword(s):  
Epithelial Cells ◽  
Microscopic Analysis ◽  
Serum Samples ◽  
Electron Microscopic ◽  
Protein Preparation ◽  
Protein Mixture ◽  
Mouse Keratinocytes ◽  
And Function ◽  
Structure Assembly

Treatment of bovine tongue mucosa with 1 M KCl induced a split in the lamina densa of the basement membrane zone (BMZ). The epithelium was then separated from the underlying connective tissue. Electron microscopic analysis of the stripped epithelium revealed that hemidesmosomes and their associated intermediate filaments (IF) remain along the basal surface of the epithelium. This surface was solubilized in an SDS/urea-containing buffer. Characterization of components of this protein mixture was undertaken using human autoantibodies from bullous pemphigoid (BP) patients that have been shown to recognize hemidesmosomal plaque elements (Mutasim, D. F., Y. Takahashi, R. S. Labib, G. J. Anhalt, H. P. Patel, and L. A. Diaz. 1985. J. Invest. Dermatol. 84:47-53) and by production of mAbs. Affinity-purified autoantibodies directed against 180- and 240-kD polypeptides present in the protein preparation generated strong immunofluorescence staining patterns along the BMZ of bovine tongue mucosa. Furthermore, immunogold localization revealed that these two polypeptides are associated with the hemidesmosomal plaque. A mAb preparation directed against a 125-kD polypeptide present in this same protein mixture lamina lucida side of the hemidesmosome. Autoantibodies in BP serum samples, affinity-purified 180-kD autoantibodies and the mAb preparation generated a punctate stain along the substratum attached surface of epithelial cells maintained on glass substrata for approximately 1 wk. The spots appeared to be associated with bundles of IF in cultured mouse keratinocytes. These monospecific antibody probes should prove invaluable for the study of hemidesmosome structure, assembly, and function.

Preparatory Procedures for Electron Microscopic Analysis at the Molecular Level of Resolution

1978 ◽  
Vol 36 (3) ◽  
pp. 516-520
Author(s):  
F.J. Sjostrand
Keyword(s):  
Electron Microscope ◽  
Molecular Level ◽  
Microscopic Analysis ◽  
Resolving Power ◽  
Cellular Structures ◽  
Electron Microscopic ◽  
Systematic Analysis ◽  
Technical Developments ◽  
Thin Sectioning ◽  
Obvious Reason

In the 1940's and 1950's electron microscopy conferences were attended with everybody interested in learning about the latest technical developments for one very obvious reason. There was the electron microscope with its outstanding performance but nobody could make very much use of it because we were lacking proper techniques to prepare biological specimens. The development of the thin sectioning technique with its perfectioning in 1952 changed the situation and systematic analysis of the structure of cells could now be pursued. Since then electron microscopists have in general become satisfied with the level of resolution at which cellular structures can be analyzed when applying this technique. There has been little interest in trying to push the limit of resolution closer to that determined by the resolving power of the electron microscope.

The Effect of Oxidized Cholesterol on the Aorta of Rabbits- Scanning Electron Microscopic Observations

1982 ◽  
Vol 40 ◽  
pp. 340-341
Author(s):  
S. K. Pena ◽  
C. B. Taylor ◽  
J. Hill ◽  
J. Safarik
Keyword(s):  
Cholesterol Biosynthesis ◽  
Cholesterol Content ◽  
Arterial Injury ◽  
Electron Microscopic ◽  
Aortic Smooth Muscle ◽  
Oxidized Cholesterol ◽  
Initial Event ◽  
Membrane Structure And Function ◽  
Scanning Electron Microscopic ◽  
And Function

Introduction: Oxidized cholesterol derivatives have been demonstrated in various cell cultures to be very potent inhibitors of 3-hvdroxy-3- methylglutaryl Coenzyme A reductase which is a principle regulator of cholesterol biosynthesis in the cell. The cholesterol content in the cells exposed to oxidized cholesterol was found to be markedly decreased. In aortic smooth muscle cells, the potency of this effect was closely related to the cytotoxicity of each derivative. Furthermore, due to the similarity of their molecular structure to that of cholesterol, these oxidized cholesterol derivatives might insert themselves into the cell membrane, alter membrane structure and function and eventually cause cell death. Arterial injury has been shown to be the initial event of atherosclerosis.

Selection and Preparation of Specific Tissue Regions For Tem Using Large Epoxy-Embedded Blocks

1973 ◽  
Vol 31 ◽  
pp. 324-325 ◽  
Author(s):  
P. M. Lowrie ◽  
W. S. Tyler
Keyword(s):  
Electron Microscopy ◽  
Anatomical Structures ◽  
Ultrastructural Studies ◽  
Transmission Electron ◽  
Microscopic Evaluation ◽  
Embedded Blocks ◽  
Specific Tissue ◽  
Definition Of ◽  
Areas Of Interest ◽  
Selection Of

The importance of examining stained 1 to 2μ plastic sections by light microscopy has long been recognized, both for increased definition of many histologic features and for selection of specimen samples to be used in ultrastructural studies. Selection of specimens with specific orien ation relative to anatomical structures becomes of critical importance in ultrastructural investigations of organs such as the lung. The uantity of blocks necessary to locate special areas of interest by random sampling is large, however, and the method is lacking in precision. Several methods have been described for selection of specific areas for electron microscopy using light microscopic evaluation of paraffin, epoxy-infiltrated, or epoxy-embedded large blocks from which thick sections were cut. Selected areas from these thick sections were subsequently removed and re-embedded or attached to blank precasted blocks and resectioned for transmission electron microscopy (TEM).

The Scanning Electron Microscopic Observation of the Vestibular Sensory Epithelia

1969 ◽  
Vol 27 ◽  
pp. 40-41
Author(s):  
D.J. Lim ◽  
W.C. Lane
Keyword(s):  
Semicircular Canals ◽  
Electron Microscopic Observation ◽  
Gravitational Acceleration ◽  
Electron Microscopic ◽  
Sensory Epithelia ◽  
Scanning Electron Microscopic ◽  
Vestibular Sensory Epithelia ◽  
And Function ◽  
Sand Piles ◽  
Active Investigation

The morphology and function of the vestibular sensory organs has been extensively studied during the last decade with the advent of electron microscopy and electrophysiology. The opening of the space age also accelerated active investigation in this area, since this organ is responsible for the sensation of balance and of linear, angular and gravitational acceleration.The vestibular sense organs are formed by the saccule, utricle and three ampullae of the semicircular canals. The maculae (sacculi and utriculi) have otolithic membranes on the top of the sensory epithelia. The otolithic membrane is formed by a layer of thick gelatin and sand-piles of calcium carbonate crystals (Fig.l).

Structural Role of rRNAs on the Ribosomal Subunits from E. Coli by Scanning Transmission Electron Microscopy

1981 ◽  
Vol 39 ◽  
pp. 424-425
Author(s):  
M. Boublik ◽  
N. Robakis ◽  
J.S. Wall
Keyword(s):  
Three Dimensional ◽  
Uranyl Acetate ◽  
Dimensional Structure ◽  
Electron Microscopic ◽  
Ribosomal Subunits ◽  
E Coli ◽  
Freeze Dried ◽  
16S Rna ◽  
Scanning Transmission ◽  
And Function

The three-dimensional structure and function of biological supramolecular complexes are, in general, determined and stabilized by conformation and interactions of their macromolecular components. In the case of ribosomes, it has been suggested that one of the functions of ribosomal RNAs is to act as a scaffold maintaining the shape of the ribosomal subunits. In order to investigate this question, we have conducted a comparative TEM and STEM study of the structure of the small 30S subunit of E. coli and its 16S RNA.The conventional electron microscopic imaging of nucleic acids is performed by spreading them in the presence of protein or detergent; the particles are contrasted by electron dense solution (uranyl acetate) or by shadowing with metal (tungsten). By using the STEM on freeze-dried specimens we have avoided the shearing forces of the spreading, and minimized both the collapse of rRNA due to air drying and the loss of resolution due to staining or shadowing. Figure 1, is a conventional (TEM) electron micrograph of 30S E. coli subunits contrasted with uranyl acetate.

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