scholarly journals Immunogold probes for light and electron microscopic localization of Ole e I in several Oleaceae pollens.

1996 ◽  
Vol 44 (2) ◽  
pp. 151-158 ◽  
Author(s):  
M C Fernandez ◽  
A Olmedilla ◽  
J D Alche ◽  
P Palomino ◽  
C Lahoz ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Major Allergen ◽  
Rapid Screening ◽  
Silver Enhancement ◽  
Electron Microscopic ◽  
Enhancement Technique ◽  
Transmission Electron ◽  
Forsythia Suspensa ◽  
Olive Pollen ◽  
Electron Microscopic Localization

We investigated the immunolocalization of the olive major allergen Ole e I and Ole e I-like proteins in pollen from several Oleaceae species [olive (Olea europaea), ash (Fraxinus excelsior), privet (Ligustrum vulgaris), lilac (Syringa vulgare), and forsythia (Forsythia suspensa)]. Crossreactions among different pollens were found in enzyme immunoassays. For immunolocalization with light microscopy we used the silver enhancement technique with three monoclonal antibodies (1D8, 10H1, and 16G2) that recognize three different epitopes of the allergen Ole e I. Our findings show that the silver enhancement technique is very useful when several antibodies are to be used for rapid screening of different materials. MAb 10H1 gave the most precise results and was selected for further immunolocalization studies with transmission electron microscopy. The epitope recognized by this MAb was localized exclusively in the endoplasmic reticulum in olive pollen. In lilac, privet, and ash pollen, most of the reactivity was also seen in the endoplasmic reticulum; however, the 10H1 epitope was not detected in forsythia pollen.

Use of Uranium-Labeled Antibodies for Electron Microscopic Localization of Various Antigens in Mammalian Cells

1967 ◽  
Vol 25 ◽  
pp. 136-137
Author(s):  
J. P. Petrali ◽  
E. J. Donati ◽  
L. A. Sternberger
Keyword(s):  
Endoplasmic Reticulum ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Specific Antiserum ◽  
Electron Microscopic ◽  
E Coli ◽  
Cytoplasmic Membranes ◽  
Viral Progeny ◽  
Ultrathin Sections ◽  
Electron Microscopic Localization

Specific contrast is conferred to subcellular antigen by applying purified antibodies, exhaustively labeled with uranium under immunospecific protection, to ultrathin sections. Use of Seligman’s principle of bridging osmium to metal via thiocarbohydrazide (TCH) intensifies specific contrast. Ultrathin sections of osmium-fixed materials were stained on the grid by application of 1) thiosemicarbazide (TSC), 2) unlabeled specific antiserum, 3) uranium-labeled anti-antibody and 4) TCH followed by reosmication. Antigens to be localized consisted of vaccinia antigen in infected HeLa cells, lysozyme in monocytes of patients with monocytic or monomyelocytic leukemia, and fibrinogen in the platelets of these leukemic patients. Control sections were stained with non-specific antiserum (E. coli).In the vaccinia-HeLa system, antigen was localized from 1 to 3 hours following infection, and was confined to degrading virus, the inner walls of numerous organelles, and other structures in cytoplasmic foci. Surrounding architecture and cellular mitochondria were unstained. 8 to 14 hours after infection, antigen was localized on the outer walls of the viral progeny, on cytoplasmic membranes, and free in the cytoplasm. Staining of endoplasmic reticulum was intense and focal early, and weak and diffuse late in infection.

scholarly journals Electron Microscopic Localization of Russell Bodies in the Human Plasma Cell

Blood ◽  
1960 ◽  
Vol 16 (3) ◽  
pp. 1307-1312 ◽  
Author(s):  
RONALD A. WELSH
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Human Plasma ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Electron Microscopic ◽  
Degenerative Process ◽  
Microscopic Localization ◽  
Electron Microscopic Localization

Abstract The location of Russell bodies in the human plasma cell was shown by electron microscopy to be within the intracisternal space of the endoplasmic reticulum. The significance of this finding was discussed from the standpoint of possible intracellular function of the endoplasmic reticulum. The appearance of the affected plasma cells tended to negate a degenerative process, and the suggestion was offered that the Russell body results from a condensation of intracisternal secretion.

A comparative light and electron microscopic study of microspore and tapetal development in male fertile and cytoplasmic male sterile oilseed rape (Brassica napus)

1986 ◽  
Vol 64 (5) ◽  
pp. 1055-1068 ◽  
Author(s):  
I. Grant ◽  
W. D. Beversdorf ◽  
R. L. Peterson
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Brassica Napus ◽  
Microspore Development ◽  
Male Sterile ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Mother Cells ◽  
Scanning Electron

The cytological development of male cells and the tapetum of male fertile and combined cytoplasmic triazine-resistant cyto-plasmic-genetic male sterile (ctr) lines of B. napus L. was studied using light, scanning electron, and transmission electron microscopy. Development of the cytoplasmic-genetic male sterile anther was similar to the normal anther up to and including meiotic prophase I. After this stage, degeneration of the microspore mother cells occurs within the callose walls, and tetrads of microspores are not formed. These degenerating microspore mother cells appear to develop numerous endoplasmic reticulum derived vesiculated structures, which may be involved in lysis of organelles. Degeneration occurs simultaneously with a proliferation of the tapetum, which eventually fills the anther locule. It is not clear whether the abortion of the microspore mother cells during meiosis stimulates proliferation of the tapetum or whether the proliferating tapetum actually interferes with microspore development thereby causing degeneration. Dilated endoplasmic reticulum cisternae containing crystalline-like deposits, and plastids with osmiophilic bodies, are frequent in cells of the proliferated tapetum of cytoplasmic-genetic male sterile anthers.

Electron microscopic localization of sulfated glycosaminoglycansans and proteoglycans in chordoma

1987 ◽  
Vol 45 ◽  
pp. 848-849
Author(s):  
Ronald Lam ◽  
Mary Ellen McGowan
Keyword(s):  
Electron Microscopy ◽  
Extracellular Matrix ◽  
Endoplasmic Reticulum ◽  
Rough Endoplasmic Reticulum ◽  
Electron Microscopic Examination ◽  
Ruthenium Red ◽  
Electron Microscopic ◽  
Cytoplasmic Vacuoles ◽  
Microscopic Studies ◽  
Electron Microscopic Localization

Although several electron microscopic studies of chordoma have been published, the origin of the chondroitin sulfate rich extracellular matrix is still not clear. Based on the ultrastructural similarities between the extracellular matrix, contents of the rough endoplasmic reticulum and cytoplasmic vacuoles, some authors assume that the two latter structures of chordoma cells contain mucosubstance. The intent of this study is to localize the sulfated glycosaminoglycans intracellularly in a chordoma, which provides cytochemical evidence of the origin of extracellular matrix.A sacrococcygeal chordoma from a 65 year old man was examined by electron microscopy after fixation in 2.5% gluta-raldehyde, 0.2% ruthenium red-glutaraldehyde, and pre-embedment staining with high iron diamine (HID), a method specific for sulfated glycoconjugates. Routine electron microscopic examination revealed stellate nonvacuolated and vacuolated “physaliferous” cells embedded in an abundant extracellular matrix. In general the chordoma cells possessed prominent Golgi complex, rough endoplasmic reticulum, mitochondria, glycogen and intermediate filaments.

Visualization of heavy metals in cultured macrophages and hepatocytes

1990 ◽  
Vol 48 (3) ◽  
pp. 960-960
Author(s):  
Ulf T Brunk ◽  
Anders Brunmark ◽  
Johann M. Zdolsek
Keyword(s):  
Heavy Metals ◽  
Cultured Cells ◽  
Electron Microscopic Level ◽  
Metal Sulfides ◽  
Silver Enhancement ◽  
Electron Microscopic ◽  
Enhancement Technique ◽  
High Ph ◽  
Organic Complexes ◽  
Sulfide Ions

The objective of this study was to develop a cytochemical method, based on a modification of the Timm method, or the silver enhancement technique, whereby iron, which is not bound within very stable metallo-organic complexes can be visualized in glutaraldehyde-fixed cultured cells with preserved ultrastructure. This has previously not been possible due to a severe damage to the fine structure caused by exposure to high pH during the sulfidation process. However, if cells are wellstabilized by glutaraldehyde before sulfidation at pH-9, they withstand the alkaline environment needed to produce a level of sulfide ions adequate to generate sufficient amounts of heavy metal sulfides before physical development. We have compared this “high pH, high S2-” variety of the silver enhancement technique, at the light and electron microscopic level, with other EM-modifications of the technique used for the detection of easily accessible heavy metals, such as zinc.

Immuno Electron Microscopic Localization of Peroxidases in the Differentiating Xylem of Populus spp.

Holzforschung ◽  
2002 ◽  
Vol 56 (4) ◽  
pp. 355-359 ◽  
Author(s):  
Y.S. Kim ◽  
S.-G. Wi ◽  
C. Grünwald ◽  
U. Schmitt
Keyword(s):  
Electron Microscopy ◽  
Secondary Wall ◽  
Secondary Cell Wall ◽  
Polyclonal Antibodies ◽  
Middle Lamella ◽  
Immunogold Labelling ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Populus Spp ◽  
Electron Microscopic Localization

Summary Peroxidases were localized in differentiating xylem cells of Populus spp. by means of immunogold labelling in combination with transmission electron microscopy (TEM). Polyclonal antibodies raised in rabbits against horseradish peroxidase were used for these experiments. Within the cytoplasm, TEM revealed a distinct labelling of the dictyosomes, indicating that these organelles are involved in the transport of peroxidases to the plasma membrane. During the formation of primary and secondary walls, cell corner regions became distinctly labelled, the developing secondary wall layers to a much lesser degree. Significant differences in the immunogold labelling between the intercorner middle lamella and the secondary cell wall layers were not observed. According to the immunogold labelling, peroxidases are incorporated into the secondary wall more or less simultaneously with deposition of polysaccharides. Correlations between peroxidase incorporation and lignification are discussed.

scholarly journals Immunoelectron microscopic demonstration of thyroglobulin and thyroid hormones in rat thyroid gland.

1987 ◽  
Vol 35 (10) ◽  
pp. 1095-1104 ◽  
Author(s):  
P K Ring ◽  
V Johanson
Keyword(s):  
Endoplasmic Reticulum ◽  
Thyroid Gland ◽  
Thyroid Hormones ◽  
Electron Microscopic ◽  
Cell Compartments ◽  
Lr White ◽  
Microscopic Demonstration ◽  
Rat Thyroid ◽  
Rat Thyroid Gland ◽  
Electron Microscopic Localization

We have developed a post-embedding immunogold technique for electron microscopic localization and quantitation of thyroglobulin (TG), thyroxine (T4), and triiodothyronine (T3) in rat thyroid. Labeling for TG was located on rough endoplasmic reticulum, Golgi apparatus, exocytotic vesicles, luminal colloid, colloid droplets, and lysosomes, whereas labeling for thyroid hormones was located on luminal colloid, colloid droplets, and lysosomes. We tested different procedures of fixation, dehydration, embedding, polymerization, and immunoincubation to optimize ultrastructural preservation and immunolabeling. Fixation with glutaraldehyde and osmium was possible with retained antigenicity. Dehydration temperature and the choice of embedding resin were the two crucial factors for good immunolabeling. Low-temperature dehydration greatly improved immunolabeling and could be combined with embedding in the methacrylate LR White or the epoxide Agar 100 (equivalent of Epon 812) polymerized at 40-60 degrees C, as the temperature during subsequent embedding and polymerization was of little importance for the immunoreactivity. Labeling on LR White sections was always higher than on Agar 100 sections. Various etching procedures were tested without improved specific labeling. Etching with hydrochloric acid gave nonspecific labeling of certain cell compartments.

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