Ultrastructural localization of nuclear antigens during interphase in mouse 3T3 fibroblasts

1989 ◽  
Vol 67 (9) ◽  
pp. 563-574 ◽  
Author(s):  
Nathalie Chaly ◽  
Gerald St. Aubin ◽  
David L. Brown
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Ultrastructural Localization ◽  
Nuclear Pores ◽  
Thin Sections ◽  
Nuclear Ultrastructure ◽  
3T3 Fibroblasts ◽  
Nuclear Antigens ◽  
Internal Component ◽  
Immunofluorescence Labelling

Thin sections of mouse 3T3 fibroblast nuclei labelled by immunoperoxidase with anti-nuclear antibodies I1, PI1, PI2, anti-peripherin, -lamin, and -centromere have been examined in the electron microscope. Staining was compared with the corresponding immunofluorescence labelling patterns, and was correlated with nuclear ultrastructure in conventionally fixed and uranyl-lead stained samples and in unlabelled immunoperoxidase controls. Peripherin was detected at the nuclear rim in a band broader and more irregular than the lamins/lamina. The peripheral components of PI1 and PI2 appear to be localized at nuclear pores and the nuclear envelope, respectively. The internal component of PI1 staining consisted of irregular patches and strands in the nucleoplasm, closely resembling snRNP staining as reported by others. Internal PI2 labelling consisted of spots distributed apparently at random in interchromatinic regions. The spots resembled labelling by antibody I1, but were fewer and more irregular in size. Neither the PI2 nor the I1 spots were centromere associated, nor could they be correlated with specific interchromatinic structures in conventional preparations and in unlabelled controls. The results support the hypothesis that the nucleus is segregated into function-specific domains, distinguished by morphology and (or) composition from surrounding regions of the nucleus.Key words: nuclear matrix, peripherin, immunoperoxidase, electron microscopy, nuclear antigens.

scholarly journals Ultrastructural immunocytochemical localization of 5-hydroxytryptamine in gastric enterochromaffin cells.

1984 ◽  
Vol 32 (6) ◽  
pp. 661-666 ◽  
Author(s):  
R D Dey ◽  
J Hoffpauir
Keyword(s):  
Electron Microscopy ◽  
Potassium Dichromate ◽  
Picric Acid ◽  
Ultrastructural Localization ◽  
Immunocytochemical Localization ◽  
Thin Sections ◽  
Glass Slides ◽  
Ec Cells ◽  
Block Face ◽  
Immunocytochemical Method

Enterochromaffin (EC) cells in the gastrointestinal tract are known to contain 5-hydroxytryptamine (5HT). The probable ultrastructural localization of 5HT in the dense core vesicles ( DCVs ) of EC cells is based on the use of histochemical techniques, such as argentaffinity and the potassium dichromate reaction. In the present paper we describe an immunocytochemical method for specifically localizing 5HT in EC cells by electron microscopy. Pieces of mucosa from the pyloric region of the rabbit stomach were prepared for electron microscopy by fixation in 0.5% glutaraldehyde-picric acid-formaldehyde without osmication , and then embedded in LX-112. Thick sections (1 micron) were mounted on glass slides and processed for the fluorescence immunocytochemical localization of 5HT. Thin sections (60-90 nm) were mounted on formvar-coated slot grids and processed for the ultrastructural immunocytochemical localization of 5HT. Both the thick and thin sections were processed by an identical procedure, beginning with a 30-min incubation in anti-5HT antiserum diluted 1:1400, followed by an IgG-FITC-gold-labeled second antibody. Fluorescent EC cells were consistently observed in the thick sections of gastric mucosa. By carefully trimming and sectioning the adjacent block face, the identical EC cell could be identified by electron microscopy. A quantitative analysis revealed the number of gold particles in EC cells to be significantly greater over the cores of DCVs than over the non-core cytoplasm or over the nucleus. Absorption of the primary antiserum with 5HT abolished all labeling, while absorption with a 5HT precursor, 5-hydroxytryptophan, did not significantly reduce core labeling. Non-EC epithelial cells were not labeled. These results demonstrate that immunoreactive 5HT in EC cells is stored in the cores of DCVs .

scholarly journals An Effective Method of Preparing Sections of Bacillus polymyxa Sporangia and Spores for Electron Microscopy

1960 ◽  
Vol 7 (2) ◽  
pp. 373-376 ◽  
Author(s):  
Pauline E. Holbert
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Electron Micrographs ◽  
Epoxy Resins ◽  
Bacillus Polymyxa ◽  
Thin Sections ◽  
Similar Images ◽  
Embedding Medium ◽  
First Time ◽  
Diamond Knife

Bacillus polymyxa sporangia and spores were prepared for examination in the electron microscope by methods whose critical features were apparently: judicious use of vacuum, to encourage complete penetration of the embedding medium; the use of epoxy resins as embedding media; and cutting of the thin sections with a diamond knife. Electron micrographs of material prepared in this manner exhibit undeformed sporangial sections. Some of the structures revealed have been shown before, though perhaps less distinctly; other structures are revealed here for the first time. While this single study does not pretend to elucidate all the complexities of sporulation in bacteria, these and similar images should make this possible, and some mention of the preparatory techniques that lead to them seems advisable at this time.

scholarly journals STUDIES BY ELECTRON MICROSCOPY OF THIN SECTIONS OF INFECTIOUS MYXOMATOSIS IN RABBITS

1952 ◽  
Vol 96 (4) ◽  
pp. 347-354 ◽  
Author(s):  
B. Epstein ◽  
Magdalena Reissig ◽  
E. De Robertis
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Host Cell ◽  
Internal Structure ◽  
Myxoma Virus ◽  
Intercellular Spaces ◽  
Thin Sections

Rabbits were inoculated with the C.P.M. strain of myxoma virus and the resulting subcutaneous tumors were fixed, embedded, and sectioned for observation with the electron microscope. Both round cells and the typical stellate myxomatous cells were observed in addition to changes in the collagen pattern at the intercellular spaces. The cytoplasm of the cells showed a great number of bodies of varying size and density, the largest of them having the size and other characteristics of the elementary bodies of the virus. Some of the bodies showed an internal structure, being formed by the tight clumping of small dense particles. Distribution curves of the diameter of the elementary bodies and of the smaller internal particles are presented. The morphological problems involved in the virus-host cell relationship are discussed in the case of the myxoma virus.

Electron microscope studies of the development of structure in Cheddar cheese

1974 ◽  
Vol 41 (3) ◽  
pp. 389-396 ◽  
Author(s):  
Alwyn M. Kimber ◽  
B. E. Brooker ◽  
D. G. Hobbs ◽  
J. H. Prentice
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Water Content ◽  
Cheddar Cheese ◽  
Incomplete Fusion ◽  
Casein Micelles ◽  
Thin Sections ◽  
Fat Globules ◽  
Granular Mass

SummaryThe development of structure in Cheddar cheese was followed using electron microscopy, of both thin sections and freeze-etched preparations. The casein micelles, at first separate, were seen to aggregate into a network, coalesce and finally form a granular mass. The fat globules, also separate at first, were gradually forced into clumps as a result of shrinkage of the casein network and incomplete fusion took place during ripening. Starter bacteria were seen trapped in the casein near the fat–casein interface, which was shown to be the region of highest water content in the mature cheese.

scholarly journals IDENTIFICATION OF GLYCOGEN IN ELECTRON MICROGRAPHS OF THIN TISSUE SECTIONS

1960 ◽  
Vol 8 (3) ◽  
pp. 575-589 ◽  
Author(s):  
Jean Paul Revel ◽  
Leonard Napolitano ◽  
Don W. Fawcett
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Animal Species ◽  
Short Exposure ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Microscopic Appearance ◽  
Tissue Sections ◽  
Lead Hydroxide ◽  
Electron Microscopic Appearance

The electron microscopic appearance of glycogen has been studied in the organs of several animal species. Glycogen almost always appears as roughly circular granules from 150 to 400 A in diameter. The intrinsic electron density of glycogen varies from tissue to tissue; however, treatment with lead hydroxide as described by Watson deeply stains the granules. Glycogen pellets were isolated from some of the tissues studied by centrifugation. Such pellets were shown to be glycogen by chemical and histochemical criteria. When thin sections of the pellet are examined under the electron microscope they can be seen to consist of densely packed granules similar to those found in the intact tissues. Such pellets are also stained for electron microscopy by short exposure to lead hydroxide.

scholarly journals ELECTRON MICROSCOPE AND EXPERIMENTAL INVESTIGATIONS OF THE NEUROFILAMENTOUS NETWORK IN DEITERS' NEURONS

1974 ◽  
Vol 61 (3) ◽  
pp. 701-722 ◽  
Author(s):  
J. Metuzals ◽  
W. E. Mushynski
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Interference Microscopy ◽  
Experimental Investigations ◽  
Rabbit Brain ◽  
Nuclear Pores ◽  
Isolated Cells ◽  
Thin Sections ◽  
Space Network ◽  
Neurofilamentous Network

The assembly of filamentous elements and their relations to the plasma membrane and to the nuclear pores have been studied in Deiters' neurons of rabbit brain. Electron microscopy of thin sections and of ectoplasm spread preparations have been integrated with physicochemical experiments and differential interference microscopy of freshly isolated cells. A neurofilamentous network extends as a continuous, three-dimensional, semilattice structure throughout the ectoplasm, the "plasma roads," and the perinuclear zone of the perikaryon. This space network consists of ∼90-Å wide neurofilaments arranged in fascicles which are interconnected by an exchange of neurofilaments. The neurofilaments consist of intercoiled ∼20-Å wide unit-filaments and are associated through cross-associating filaments with other neurofilaments of the fascicle and with microfilaments. The ∼20–50-Å wide microfilaments display intimate associations with the plasma membrane and with the nuclear pores. Electron microscopy of thin sections from glycerinated and heavy meromyosin-treated Deiters' neurons shows that actin-like filaments are present in the pre- and postsynaptic regions of synapses terminating on these neurons. It is proposed that the neurofilamentous space network serves a transducing function by linking plasma membrane activities with the genetic machinery of the neuron.

Modification for Electron Miscroscopy of the Corbett Ammoniacal Silver Stain

1971 ◽  
Vol 29 ◽  
pp. 484-485
Author(s):  
Robert L. Corbett
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Light Microscopy ◽  
Low Power ◽  
Light And Electron Microscopy ◽  
Glass Slide ◽  
Silver Stain ◽  
Thin Sections ◽  
Silver Deposits ◽  
Ammoniacal Silver

The ammoniacal silver stain for light microscopy of plastic embedded sections has been adapted for use in electron microscopy. Since the silver will stain even very thin sections, i.e., silver and gold for light microscopy, and silver deposits are sufficiently electron dense to be seen in the electron microscope, the results are very useful for correlating light and electron microscopy. Compared to the conventional stains for electron microscopy which usually take over one-half hour, the silver procedure can be done in five minutes or less and thus provides a quick look at sections This stain has more contrast, so it is especially good for low power electron microscopy. The ability of the silver to stain very thin sections enables a correlation between light and electron microscopy in three ways. First, thin sections can be stained with silver on a glass slide and compared with immediately adjacent thin sections on grids stained the usual way for electron microscopy.

The Effect of Electron Microscopy on Pathological Diagnosis of Neoplasm

1990 ◽  
Vol 48 (3) ◽  
pp. 248-249
Author(s):  
Xiaojun Zhou ◽  
Taihe Zhang
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Frozen Section ◽  
Protein A ◽  
Functional Classification ◽  
Lead Citrate ◽  
Thin Sections ◽  
Tumor Tissues ◽  
Frozen Section Diagnosis

Although electron microscopy (EM) has contributed enormously to an understanding of the structural intricacies of tumor cells, the usefulness of EM in pathological diagnoses of neoplasms has not been readily appreciated by general pathologists. In the present study, 223 tumors submitted for EM diagnosis were analyzed in an attempt to gain further information concerning the contribution of EM to tumor diagnosis.223 neoplasms were submitted to EM for their final diagnoses when histopathological diagnoses were obscure, which represented about of the total number of surgical tumor specimens. Most specimens were taken at the time of frozen section diagnosis and a small number of tissues were originally fixed informaldehyde. All of tissues were fixed with buffered glutaradehyde, postfixed with osmium tetroxide and embedded in Epon 812. Ultrasections were made after semith in sections were examined to verify that representative tumor tissues were present. Thin sections were stained with uranium acetate and lead citrate, and examined under JEM-1200 EX electron microscope. In selected cases, mainly with neuroendocrine tumors, nickel grid-mounted sections were subjected to post embedding immunoelectron microscopy (IEM) using protein A-gold for more detailed functional classification. Protein A-gold probes were prepared as Wang and co-workers described.

Nuclear pores at prophase of meiosis in plants

1974 ◽  
Vol 268 (891) ◽  
pp. 95-100 ◽  
Keyword(s):  
Fine Structure ◽  
Electron Microscope ◽  
Osmotic Shock ◽  
Nuclear Pores ◽  
Variable Size ◽  
Thin Sections ◽  
Amorphous Substance ◽  
Mother Cells ◽  
Prophase Of Meiosis

Nuclear pores have been studied with the electron microscope in thin sections of pollen mother cells at early- to mid-meiotic prophase ( a ) in respect of distribution, ( b ) in relation to fine structure in the pore complex in the following plants: Fritillaria lanceolata, Phaedranassa viridijlora, Tulbaghia violacea , an F 1 hybrid of Allium fisultosum x Allium cepa and the lily var. 'Formobel'. In all plants from leptotene to pachytene, the pores were irregularly spread over the envelope in random clusters of variable size encircled by areas in which they did not occur. Further proof was obtained from the lily for the premise that pores are not formed in regions of the envelope to which the nucleolus is adpressed at leptotene. The fine structure of the pore complex observed supports a model which proposes that annuli are composed of three rings of eight granular subunits. Most pores contained a central granule ranging from 25 to 30 nm in diameter composed of amorphous substance and filaments about 3 nm wide, apparently continuous with filaments of similar dimensions in the symmetrical annular subunits that encircle the orifice at both the nuclear and cytoplasmic sides of the pore. The pore complex and central granule were relatively more stable to osmotic shock than the ribosomal region of the nucleolus. Recent ideas concerning the role of the annulus and central granule in nucleocytoplasmic transfer of ribonucleoprotein and assembly of polyribosomes are discussed.

Spermiogenesis of Eledone cirrhosa Lamarck (Cephalopoda, Octopoda)

1974 ◽  
Vol 186 (1083) ◽  
pp. 181-190 ◽  
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Vas Deferens ◽  
Mature Male ◽  
Axial Filament ◽  
Thin Sections ◽  
Sperm Tail ◽  
Mature Gamete ◽  
Microtubular System

Spermiogenesis in a mature male Eledone cirrhosa has been studied by means of phase con­trast and electron microscopy. Material from the testis was either studied alive or by means of thin sections. Material from the vas deferens and spermatophores was either studied live, in thin sections or in negatively stained preparations for the electron microscope. The spherical spermatid cells each become modified to give an elongate mature gamete. The gamete is remarkable in the size and complexity of its spiral head, the small size of the mid-piece and the length of the sperm tail. The acrosome is simple but helical. The flagellum originates in a single centriole; the ‘proximal’ centriole is lost. The axial filament consists of a ‘normal’ 9 + 2 microtubular system together with 9 accessory γ fibres; the γ fibres form a centriolar ring at their point of origin. Cytochemical tests show that there is a loosely packed deposit of glycogen throughout the length of the tail. There are no glycogen deposits within the mid-piece or the head.

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