scholarly journals Intracellular localization of thymosin alpha 1 by immunoelectron microscopy using a monoclonal antibody.

1987 ◽  
Vol 35 (2) ◽  
pp. 181-187 ◽  
Author(s):  
C Auger ◽  
C Stahli ◽  
N Fabien ◽  
J C Monier
Keyword(s):  
Monoclonal Antibody ◽  
Epithelial Cells ◽  
Intracellular Localization ◽  
Immunofluorescence Assay ◽  
Secretory Process ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Thymic Hormone ◽  
Thin Sections ◽  
Thymosin Alpha 1

Distribution of thymosin alpha 1 in normal mice (OF1) or autoimmune mice (NZB) was investigated using immunocytochemical techniques on sections of GMA- and Epon-embedded mouse thymuses. A monoclonal antibody directed against synthetic thymosin alpha 1 was used. With the immunofluorescence assay, patchy staining of thymosin alpha 1 was found in the cytoplasm of epithelial cells of the subcapsullary and medullary zones of OF1 thymus. In NZB thymus, the fluorescent pattern was less precisely localized. At the electron microscopic level, immunolabeling of Epon-embedded ultra-thin sections revealed ferritin in some vacuoles of epithelial cells. Ferritin labeling in OF1 thymus was found in several small vacuoles of the same cell, but was present in large, dense vacuoles in NZB thymus. These differences might reflect differences in the secretory process of thymic hormone.

Correlative immunolocalization with high-resolution light microscopy and Electron Microscopy using London Resin Gold embedded tissue

1995 ◽  
Vol 53 ◽  
pp. 702-703
Author(s):  
M. L. Grove ◽  
B. A. Evans ◽  
D. N. Misra ◽  
J. Zhao ◽  
D. H. Alpers ◽  
...  
Keyword(s):  
Cellular Localization ◽  
Polyclonal Antibodies ◽  
Intracellular Localization ◽  
Intrinsic Factor ◽  
Gastric Epithelium ◽  
Electron Microscopic Level ◽  
Immunoperoxidase Staining ◽  
Rat Tissues ◽  
Electron Microscopic ◽  
Thin Sections

Immunoelectron microscopy specimens are often embedded in hydrophilic resins, like London Resin Gold(LRG), which permit antibody staining without etching (or deplasticizing) the sections. This characteristic of hydrophilic resins also allows for immunohistochemistry at the light level on semi thin sections (0.5μm - 1μm). The ability to do immunohistochemistry at both the light and electron microscopic level on the same tissue block allows focused ultrastructural study. Immunohistochemistry on semi-thin sections displays cellular localization of macromolecules, permitting more specificity in the selection of areas for studying intracellular localization ultrastructurally. We have developed a method for immunoperoxidase staining of LRG embedded tissues, utilizing anti-human polyclonal antibodies directed against intrinsic factor (Fig. 1). Intrinsic factor (IF), a cobalamin binding protein, is known to be produced in the stomach, pancreas and salivary glands of most mammals. We are interested in distribution of IF in gastric epithelium, small intestine (ileum) and supporting tissues in both gastrointestinal tract sites. Previously, we have described the cellular localization of IF in human and rat tissues.

The Immuno-Electron Microscopic Localization of the Mo-Fe Protein Component of Nitrogenase in Cells of Azotobacter Vinelandii

1973 ◽  
Vol 31 ◽  
pp. 574-575
Author(s):  
J. T. Stasny ◽  
R. C. Burns ◽  
R. W. F. Hardy
Keyword(s):  
Cellular Localization ◽  
Direct Evidence ◽  
Azotobacter Vinelandii ◽  
Intracellular Localization ◽  
Protein Component ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Fe Protein ◽  
Intracytoplasmic Membranes

Structure-functlon studies of biological N2-fixation have correlated the presence of the enzyme nitrogenase with increased numbers of intracytoplasmic membranes in Azotobacter. However no direct evidence has been provided for the internal cellular localization of any nitrogenase. Recent advances concerned with the crystallizatiorTand the electron microscopic characterization of the Mo-Fe protein component of Azotobacter nitrogenase, prompted the use of this purified protein to obtain antibodies (Ab) to be conjugated to electron dense markers for the intracellular localization of the protein by electron microscopy. The present study describes the use of ferritin conjugated to goat antitMo-Fe protein immunoglobulin (IgG) and the observations following its topical application to thin sections of N2-grown Azotobacter.

scholarly journals Application of avidin-biotin-peroxidase complex method with a monoclonal antibody to human Ia-like antigen in immunoelectron microscopy.

1986 ◽  
Vol 34 (11) ◽  
pp. 1495-1499 ◽  
Author(s):  
K C Feng-Chen ◽  
B F Chen ◽  
Z Liu ◽  
A K Ng
Keyword(s):  
Monoclonal Antibody ◽  
Ultrastructural Level ◽  
Electron Microscopic Level ◽  
Complex Method ◽  
Outer Cell ◽  
Electron Microscopic ◽  
Hla Dr ◽  
Combined Use ◽  
Antibody Complex ◽  
Reticular Cells

Monoclonal antibody (MAb) to human Ia-like (HLA-DR) antigen was applied with the avidin-biotin-peroxidase complex (ABC) immunostaining method to localize the Ia-like antigen at the electron microscopic level. Our results indicated that in human tonsils and adenoids fixed with 4-6% phosphate-buffered paraformaldehyde for 4-6 hr, sharply delineated electron-dense products of the antigen and antibody complex were detectable on the outer cell membranes of lymphoblasts, lymphocytes, reticular cells, and macrophages. In our study, the vibratome sections of the paraformaldehyde-fixed, pre-embedding immunostained tissues consistently showed more satisfactory morphology than frozen sections. The combined use of the anti-human Ia monoclonal antibody and the ABC procedure with paraformaldehyde fixation provides a simple and sensitive method to study at the ultrastructural level the Ia-like antigen-bearing cells, which are vital in the immune response.

Electron microscopic analysis of objects in light microscopic sections

1978 ◽  
Vol 36 (2) ◽  
pp. 80-81
Author(s):  
Arvid B. Maunsbach
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Microscopic Analysis ◽  
Electron Microscopic Level ◽  
Semithin Section ◽  
Cover Glass ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Glass Slides ◽  
Ultrathin Sections

Structural studies in experimental biology or in pathology are frequently extended from the light to the electron microscopic level. This is often done by cutting both semithin (about 1 μm) and thin sections from the same tissue block after embedding for electron microscopy. However, in many studies it would be of great value to analyse the same structure both by light and electron microscopy, i.e. to be able to study by electron microscopy an object which is first detected by light microscopy in a semithin section. To achieve this, a method has been developed by which ultrathin sections are cut directly from the semithin section containing the object of interest.Semithin sections, about 1 μ in thickness, are cut from Epon or Vestopal embedded tissue. The sections are placed on ordinary glass slides and stained with toluidine blue. The sections are studied in the light microscope without a cover glass or mounted in water.

scholarly journals Calmodulin immunoelectron microscopy: redistribution during ram spermatogenesis and epididymal maturation. II.

1986 ◽  
Vol 34 (9) ◽  
pp. 1181-1193 ◽  
Author(s):  
S Weinman ◽  
C Ores-Carton ◽  
F Escaig ◽  
J Feinberg ◽  
S Puszkin
Keyword(s):  
Male Gamete ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Epididymal Maturation ◽  
Coarse Fibers ◽  
Preferential Location ◽  
Lowicryl K4m ◽  
Pleiotropic Regulator ◽  
Monospecific Antibodies

Affinity-purified monospecific antibodies and indirect immunogold and immunoferritin labeling on ultra-thin sections of low-temperature Lowicryl K4M-embedded samples were used to study the redistribution of calmodulin in ram spermatids and epididymal spermatozoa at the electron microscopic level. Calmodulin appeared as an integral component of well-defined structures or organelles of these cells. In young spermatids, calmodulin was localized in the nucleus, cytoplasm, and developing acrosome. During spermatogenesis and epididymal maturation, calmodulin left the acrosome to reach the perinuclear substance and finally became concentrated in the post-acrosomal area of the head, although some calmodulin remained associated with the tip of the acrosome. Such a redistribution is consistent with the preferential location of Ca2+ in the post-acrosomal cytoplasm of ejaculated spermatozoa. Calmodulin was also observed in the flagellum associated with the plasma membrane and with the motility apparatus, between coarse fibers and axonemal microtubules. These changes in calmodulin distribution may account for the Ca2+-dependent regulation of spermatogenesis and sperm maturation. Calmodulin therefore appears to be a pleiotropic regulator of male gamete development and functions.

scholarly journals Immunohistochemical localization of acetylcholine receptors at human endplates using a monoclonal antibody.

1987 ◽  
Vol 35 (5) ◽  
pp. 613-617 ◽  
Author(s):  
L M Smit ◽  
H Veldman ◽  
F G Jennekens
Keyword(s):  
Monoclonal Antibody ◽  
Acetylcholine Receptors ◽  
Immunohistochemical Localization ◽  
Immunohistochemical Method ◽  
Electron Microscopic Level ◽  
Immunoperoxidase Technique ◽  
Electron Microscopic ◽  
Fixed Tissue ◽  
Alpha Bungarotoxin ◽  
Diagnostic Investigations

We describe a simple indirect immunohistochemical method for localization of acetylcholine receptors (AChR) in motor endplates at the light and electron microscopic level. This method involves the use of a monoclonal antibody directed against the main immunogenic region (MIR) of AChRs and is applicable to periodate-lysine-paraformaldehyde (PLP)-fixed tissue. We discuss the advantages of this method, as compared with the alpha-bungarotoxin-immunoperoxidase technique, and stress its value for diagnostic investigations of motor point biopsies from patients with neuromuscular transmission disorders.

Monoclonal antibody study of the decorated spongiome of contractile vacuole complexes of Paramecium

1990 ◽  
Vol 96 (3) ◽  
pp. 469-475
Author(s):  
R.D. Allen ◽  
M.S. Ueno ◽  
L.W. Pollard ◽  
A.K. Fok
Keyword(s):  
Monoclonal Antibody ◽  
Developmental Stages ◽  
General Scheme ◽  
Central Zone ◽  
Immunogold Labeling ◽  
Microscopic Level ◽  
Contractile Vacuole ◽  
Electron Microscopic Level ◽  
Frozen Sections ◽  
Electron Microscopic

A monoclonal antibody (mAb) has been developed and selected by immunofluorescence for the radial canals of the contractile vacuole complex (CVC) of Paramecium multimicronucleatum. By applying indirect immunogold labeling to thin frozen sections this mAb has been shown at the electron microscopic level to be specific for the decorated spongiome. We have used the mAb to study the normal interfission appearance as well as developmental stages of the decorated spongiomes. Two decorated spongiomes, presumably involved in water sequestration, radiate as 5–10 bands from unlabeled, circular, 25 microns diameter centers. Two new CVCs arise just anterior to the space occupied by the old spongiomes, the new anterior CVC appearing slightly before the posterior one. Development of the new spongiomes around a 10 microns unlabeled central zone is accompanied by a regression of old spongiome bands until the lengths of these bands in both old and new CVCs are equal just before cell division. After division both old and new spongiome bands grow at equal rates to the same length. Exceptions to the above general scheme, both in number of CVCs in interfission, as well as in position of the new relative to the old CVCs, are also observed.

Cell-to-cell adherens junction formation and actin filament organization: similarities and differences between non-polarized fibroblasts and polarized epithelial cells

1995 ◽  
Vol 108 (1) ◽  
pp. 127-142 ◽  
Author(s):  
S. Yonemura ◽  
M. Itoh ◽  
A. Nagafuchi ◽  
S. Tsukita
Keyword(s):  
Epithelial Cells ◽  
Early Stage ◽  
Light And Electron Microscopy ◽  
Adherens Junction ◽  
Cell Polarization ◽  
Stress Fiber ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Similarities And Differences ◽  
Polarized Epithelial Cells

Cadherin has an intimate spatial relationship with actin filaments (AF) in various types of cells, forming the cell-to-cell adherens junction (AJ). We compared the AJ/AF relationship between non-polarized fibroblasts (NRK cells) and polarized epithelial cells (MTD-1A cells). E/P-cadherin, alpha-catenin, ZO-1 and vinculin were localized with reference to AF in these cells using laser scan microscopy as well as conventional light and electron microscopy. NRK cells adhered to each other at the tips of thin cellular processes, where spot-like AJ were formed, where P-cadherin, alpha-catenin, ZO-1 and vinculin were concentrated. Some stress-fiber-like AF bundles ran axially in these processes and terminated at spot-like AJ on their tips. At the electron microscopic level these spot-like AJ were seen as aggregates of small ‘units’ of AJ, where AF were densely and perpendicularly associated with the plasma membrane. In MTD-1A cells, the AJ/AF relationship was investigated during the cell polarization process after replating or wounding. At the early stage, the AJ/AF relationship was quite similar to that in NRK cells. As polarization proceeded, the spot-like AJs were gradually fused side by side with the concomitant shortening of the associated stress-fiber-like AF bundles. Finally, the belt-like AJ was established, which was lined with circumferential AF bundles. The similarities and differences in the AJ/AF relationship between non-polarized fibroblasts and polarized epithelial cells are discussed.

Morphologic alterations in the parietal yolk-sac of the rat from the 12th to the 19th day of gestation

Development ◽  
1977 ◽  
Vol 39 (1) ◽  
pp. 9-21
Author(s):  
R. P. Jensh ◽  
T. R. Koszalka ◽  
M. Jensen ◽  
L. Biddle ◽  
R. L. Brent
Keyword(s):  
Epithelial Cells ◽  
Experimental Models ◽  
Yolk Sac ◽  
Basement Membranes ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
En Face ◽  
Pas Staining ◽  
Reichert's Membrane ◽  
Histologic Changes

The rat parietal yolk-sac and its adherent epithelial cells were examined at various stages of gestation using an en face technique. Specimens were observed at both the light and electron microscopic level. Diastase pretreatment and PAS-staining were used to determine the presence of glycogen. As early as the 12th day of gestation the cytoplasm of the parietal yolk-sac cells contained numerous ribosomes and mitochondria and a large amount of endoplasmic reticulum. The glycogen content of the epithelial cells increased from the 12th day of gestation and accumulated in large quantities by the 16th day. By the 17th day many cells exhibited variable degrees of degeneration. Cellular elements of degenerating cells appeared to be trapped within Reichert's membrane. Contrary to the reports of other investigators, the present study indicates that the capsular portion of the parietal yolk-sac consisting of Reichert's membrane and its adherent epithelial cells remained intact until at least the 18th day of gestation. Some of the unique characteristics of the parietal yolk-sac provide experimental models to study the effects of environmental factors on (1) the synthesis of basement membranes, (2) the ageing of cells and (3) the correlation of these histologic changes with the functions of the parietal yolk-sac.

scholarly journals Distribution of the slow/cardiac isoform of skeletal muscle Ca(2+)-ATPase in developing and mature tissues of chickens determined using a monoclonal antibody.

1993 ◽  
Vol 41 (2) ◽  
pp. 215-224
Author(s):  
S Shahin ◽  
P F Bartlett ◽  
T J Millar ◽  
I McLennan ◽  
J A Rostas
Keyword(s):  
Skeletal Muscle ◽  
Monoclonal Antibody ◽  
Smooth Muscle ◽  
Muscle Fibers ◽  
Cell Types ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Adult Chicken ◽  
Intrafusal Muscle Fibers ◽  
Anterior Latissimus Dorsi

We have established that the monoclonal antibody (MAb) AA21, raised against a crude sarcolemmal fraction prepared from adult chicken anterior latissimus dorsi muscle, recognizes the slow twitch/cardiac isoform of calcium ATPase. This was done using a combination of immunohistochemistry at the light and electron microscopic level, the change in the cell distribution in skeletal muscle during development, the molecular weight of the principal protein recognized in Western transfers, and direct comparison with another MAb of known specificity. The antigen is initially expressed by all myotubes at E10 and with development is gradually lost from all presumptive fast fibers. In addition to its immunoreaction and slow extrafusal skeletal muscle fibers, AA21 displays a highly selective immunoreactivity with a number of other cell types in different tissues. The antibody stains a subset of intrafusal muscle fibers and intestinal and arterial smooth muscle, but not venous smooth muscle. In the nervous system, a subpopulation of neurons is intensely stained, most neurons are faintly stained, and glia are not stained at all.

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