A set of cell surface glycoproteins forms an early position, but not cell type, in the root apical carota L

Development ◽  
1989 ◽  
Vol 106 (1) ◽  
pp. 47-56
Author(s):  
J.P. Knox ◽  
S. Day ◽  
K. Roberts
Keyword(s):  
Monoclonal Antibody ◽  
Vascular Tissue ◽  
Cultured Cells ◽  
Frozen Sections ◽  
Cell Type ◽  
Daucus Carota L ◽  
Seedling Root ◽  
Carrot Cell ◽  
Cell Surface Glycoproteins ◽  
Ultrathin Frozen Sections

A monoclonal antibody (JIM4) has been derived that series of glycoproteins associated with the plasma suspension-cultured carrot cell line and also an proteoglycan secreted by the cultured cells. indicated that the plasma membrane antigen(s) specific to certain cells of Daucus carota L. apex JIM4 labelled two segments of the stele. These the poles of the protoxylem. An axis of unlabelled two phloem regions. Two sections of the pericycle with oblique longitudinal divisions were particularly This pattern of reactive cells, reflecting cell specific future cell type, would appear to be a unique plants. The association of JIM4 antigens with these maintained through the transition from root to the cotyledons and the mature plant. Examination of JIM4 ultrathin frozen sections of the carrot seedling root indicated that the expression of the antigen is a very root development. Cells express the surface epitope, cells of the dome of apical initials, before the vascular tissue can be discerned and well before its differentiation.

FUSION OF CARROT AND BARLEY PROTOPLASTS AND DIVISION OF HETEROKARYOCYTES

1976 ◽  
Vol 18 (2) ◽  
pp. 263-269 ◽  
Author(s):  
D. Dudits ◽  
K. N. Kao ◽  
F. Constabel ◽  
O. L. Gamborg
Keyword(s):  
Cell Walls ◽  
Culture Medium ◽  
Cultured Cells ◽  
Differential Staining ◽  
Hordeum Vulgare L ◽  
Daucus Carota L ◽  
Carrot Cell ◽  
Fusion Of Protoplasts ◽  
Fusion Products ◽  
Carrot Cell Cultures

Fusion of protoplasts from cultured cells of carrot (Daucus carota L.) and from leaves of barley (Hordeum vulgare L.) by means of polyethylene glycol resulted in the formation of 4-5 fusion products (heterokaryocytes) per 100 protoplasts. When incubated in culture medium, the heterokaryocytes regenerated cell walls and divided. The frequency of division depended on the viability of the protoplasts from carrot cell cultures, specifically, on the mitotic activity of the cells. Fusion of interphase carrot and barley nuclei was detected by differential staining. Synchronized mitosis was observed in heterokaryons containing barley and carrot nuclei.

scholarly journals ULTRATHIN FROZEN SECTIONS

1967 ◽  
Vol 34 (3) ◽  
pp. 757-771 ◽  
Author(s):  
W. Bernhard ◽  
Elizabeth H. Leduc
Keyword(s):  
Electron Microscopy ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Phosphotungstic Acid ◽  
Cultured Cells ◽  
Uranyl Acetate ◽  
Distilled Water ◽  
Frozen Sections ◽  
Simple Method ◽  
Ultrathin Frozen Sections

A relatively simple method for obtaining ultrathin, frozen sections for electron microscopy has been developed. Tissues, cultured cells, and bacteria may be employed. They are fixed in 1.25–4% glutaraldehyde for 1–4 hr, are washed overnight in buffer at 3°C, and are embedded in 20% thiolated gelatin or pure gelatin. Before sectioning they are partially dehydrated in 50% glycerol, frozen in liquid nitrogen on a modified tissue holder, and subsequently maintained at -70°C with dry ice. Finally, they are sectioned very rapidly with glass knives on a slightly modified Porter-Blum MT-1 microtome in a commercial deep-freeze maintained at -35°C and are floated in the trough of the knife on a 40% solution of dimethylsulfoxide (DMSO). The sections are picked up in plastic loops and transferred to distilled water at room temperature for thawing and removal of the DMSO, placed on grids coated with Formvar and carbon, air-dried, and stained with phosphotungstic acid, sodium silicotungstate, or a triple stain of osmium tetroxide, uranyl acetate, and lead. Large flat sections are obtained in which ultrastructural preservation is good. They are particularly useful for cytochemical studies.

Cingulin: characterization and localization

1989 ◽  
Vol 93 (1) ◽  
pp. 107-122 ◽  
Author(s):  
S. Citi ◽  
H. Sabanay ◽  
J. Kendrick-Jones ◽  
B. Geiger
Keyword(s):  
Tight Junction ◽  
Cultured Cells ◽  
Stress Fibers ◽  
Junctional Complex ◽  
Kidney Cells ◽  
Frozen Sections ◽  
Heat Stable ◽  
Polarized Epithelia ◽  
Stokes Radius ◽  
Ultrathin Frozen Sections

Cingulin, a protein component associated with the tight junctions of chicken intestinal epithelium, has been purified to homogeneity by a new procedure and characterized. Purified cingulin is a heat-stable elongated dimer, composed of two polypeptides of Mr 108,000 (cingulin-108), with a Stokes' radius of approximately 15 nm, and a molecular length of 130 nm +/− 32 nm. Monoclonal antibodies were used to determine the tissue distribution and subcellular localization of cingulin in a variety of avian tissues and cultured cells. Indirect immunofluorescence analysis of semi-thin frozen sections demonstrated that cingulin is localized in the junctional complex of various polarized epithelia and in the endothelium, whereas it is essentially absent from mesenchymal and myogenic cells. In permeabilized and fixed cultured chick embryo kidney cells, the antibodies stained solely the regions of contacts between the epithelial cells. Double immunofluorescent labeling of these cells with anti-cingulin and anti-vinculin antibodies showed that cingulin is localized close to the vinculin-rich cytoskeletal belt associated with adherens junctions, but is absent from focal contacts and stress fibers. In cultured kidney cells, actin was detected mainly in stress fibers and in the peripheral junctional regions, where it showed a distribution similar to that of cingulin, suggesting that actin filaments may be part of the submembrane cytoskeleton at the level of the tight junction. Indirect immunoelectron microscopic labeling of ultrathin frozen sections of chicken intestine showed that cingulin is localized along the endofacial surfaces of the tight junction (zonula occludens), and is apparently excluded from the more basal zonula adhaerens, and from the desmosomes.

Immunoferritin localization of intracellular antigens in positively stained frozen sections

1978 ◽  
Vol 36 (2) ◽  
pp. 168-169
Author(s):  
K. T. Tokuyasu
Keyword(s):  
Surface Tension ◽  
Water Surface ◽  
Thin Layers ◽  
The Other ◽  
Positive Staining ◽  
Frozen Sections ◽  
The Past ◽  
Ultrathin Frozen Sections ◽  
Definition Of

During the past investigations of immunoferritin localization of intracellular antigens in ultrathin frozen sections, we found that the degree of negative staining required to delineate u1trastructural details was often too dense for the recognition of ferritin particles. The quality of positive staining of ultrathin frozen sections, on the other hand, has generally been far inferior to that attainable in conventional plastic embedded sections, particularly in the definition of membranes. As we discussed before, a main cause of this difficulty seemed to be the vulnerability of frozen sections to the damaging effects of air-water surface tension at the time of drying of the sections.Indeed, we found that the quality of positive staining is greatly improved when positively stained frozen sections are protected against the effects of surface tension by embedding them in thin layers of mechanically stable materials at the time of drying (unpublished).

Localization of pancreatic enzymes in ultrathin frozen sections stained with metal labeled antibody

1978 ◽  
Vol 36 (2) ◽  
pp. 90-91
Author(s):  
Kenjiro Yasuda
Keyword(s):  
Fluorescent Antibody ◽  
Pancreatic Enzymes ◽  
Tissue Preparation ◽  
Zymogen Granules ◽  
Frozen Sections ◽  
En Bloc ◽  
Antibody Method ◽  
Ultrathin Frozen Sections ◽  
Fluorescent Antibody Method

Localization of amylase,chymotrypsinogen and trypsinogen in pancreas was demonstrated by Yasuda and Coons (1966), by using fluorescent antibody method. These enzymes were naturally found in the zymogen granules. Among them, amylase showed a diffuse localization around the nucleus, in addition to the zymogen granules. Using ferritin antibody method, scattered ferritin granules were also found around the Golgi area (Yasuda et al.,1967). The recent advance in the tissue preparation enables the antigen to be localized in the ultrathin frozen sections, by applying the labeled antibodies onto the sections instead of staining the tissue en bloc.The present study deals with the comparison of the localization of amylase and lipase demonstrated by applying the bismuth-labeled, peroxidase-labeled and ferritin-labeled antibody methods on the ultrathin frozen sections of pancreas, and on the blocks of the same tissue.

The Application of Ultracryotomy to Immunocytochemistry

1973 ◽  
Vol 31 ◽  
pp. 704-709
Author(s):  
R. G. Painter ◽  
K. T. Tokuyasu ◽  
S. J. Singer
Keyword(s):  
Frozen Sections ◽  
Protein Antigens ◽  
Carbon Coated ◽  
Antibody Conjugates ◽  
Ultrathin Frozen Sections

A technique for localizing intracellular antigens with immunoferritin conjugates directly on ultrathin frozen sections of glutaraldehyde-fixed tissues has been developed. This method overcomes some of the limitations of previously described procedures, since it avoids drastic fixation, dehydration and embedding procedures which could denature many protein antigens.Briefly cells or tissues were fixed with glutaraldehyde (0.5 to 2% for 1 hr), and ultrathin frozen sections were cut and mounted on grids covered with carbon-coated Formvar film by the procedure described previously. Such sections were stained with ferritin-antibody conjugates by methods described elsewhere.

Ultrathin frozen sections of yeast without aldehyde prefixation

1978 ◽  
Vol 36 (2) ◽  
pp. 58-59
Author(s):  
K. J. Böhm ◽  
a. E. Unger
Keyword(s):  
Aqueous Solutions ◽  
Biological Material ◽  
Yeast Cells ◽  
Frozen Sections ◽  
Good Preservation ◽  
Ultrathin Frozen Sections

During the last years it was shown that also by means of cryo-ultra-microtomy a good preservation of substructural details of biological material was possible. However the specimen generally was prefixed in these cases with aldehydes.Preparing ultrathin frozen sections of chemically non-prefixed material commonly was linked up to considerable technical and manual expense and the results were not always satisfying. Furthermore, it seems to be impossible to carry out cytochemical investigations by means of treating sections of unfixed biological material with aqueous solutions.We therefore tried to overcome these difficulties by preparing yeast cells (S. cerevisiae) in the following manner:

Ultrastructural Correlations between Clonal Human Tumor Colonies and in Vivo Neoplasms

1982 ◽  
Vol 40 ◽  
pp. 210-211
Author(s):  
M.J. Murphy ◽  
R.R. Price ◽  
J.C. Sloman
Keyword(s):  
Cultured Cells ◽  
Human Tumor ◽  
Cell Type ◽  
Tumor Mass ◽  
Initial Cell ◽  
Cloning Assay ◽  
Human Tumor Cloning ◽  
The Relationship

The in vitro human tumor cloning assay originally described by Salmon and Hamburger has been applied recently to the investigation of differential anti-tumor drug sensitivities over a broad range of human neoplasms. A major problem in the acceptance of this technique has been the question of the relationship between the cultured cells and the original patient tumor, i.e., whether the colonies that develop derive from the neoplasm or from some other cell type within the initial cell population. A study of the ultrastructural morphology of the cultured cells vs. patient tumor has therefore been undertaken to resolve this question. Direct correlation was assured by division of a common tumor mass at surgical resection, one biopsy being fixed for TEM studies, the second being rapidly transported to the laboratory for culture.

scholarly journals Assembly of vimentin in cultured cells varies with cell type

1989 ◽  
Vol 264 (30) ◽  
pp. 17953-17960
Author(s):  
W B Isaacs ◽  
R K Cook ◽  
J C Van Atta ◽  
C M Redmond ◽  
A B Fulton
Keyword(s):  
Cultured Cells ◽  
Cell Type
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