The subcellular localization of immunoglobulin in mouse plasma cells, as studied with immunoferritin cytochemistry on ultrathin frozen sections

1980 ◽  
Vol 158 (2) ◽  
pp. 161-169 ◽  
Author(s):  
Hans J. Geuze ◽  
Jan W. Slot
Keyword(s):  
Subcellular Localization ◽  
Plasma Cells ◽  
Frozen Sections ◽  
Mouse Plasma ◽  
Ultrathin Frozen Sections

scholarly journals ULTRASTRUCTURAL LOCALIZATION OF INTRACELLULAR IMMUNE GLOBULINS IN PLASMA CELLS AND LYMPHOBLASTS BY ENZYME-LABELED ANTIBODIES

1969 ◽  
Vol 17 (4) ◽  
pp. 211-224 ◽  
Author(s):  
ELIZABETH H. LEDUC ◽  
GEOFFREY B. SCOTT ◽  
STRATIS AVRAMEAS
Keyword(s):  
Immune Response ◽  
Plasma Cells ◽  
Ultrastructural Localization ◽  
Frozen Sections ◽  
Cytoplasmic Staining ◽  
Specific Antibodies ◽  
Perinuclear Space ◽  
Cacodylate Buffer ◽  
Immune Globulins ◽  
Ultrathin Frozen Sections

Techniques for localization of immune globulins by specific antibodies labeled with peroxidase or alkaline phosphatase were applied to spleens of rabbits hyperimmunized against various antigens. Best results were obtained with tissue blocks or thick sections after fixation in 1% formaldehyde (freshly prepared from paraformaldehyde) in cacodylate buffer with sucrose. The edges of blocks gave adequate penetration, comparable to thick sections. Ultrathin frozen sections which were reacted directly with labeled conjugate were less successful and glycol methacrylate sections failed to stain. Results were 3-fold: (1) immune globulins were demonstrated in ergastoplasmic cisternae, perinuclear space and Golgi apparatus of plasmocytes; (2) associated ribosomal staining varied somewhat with fixation and osmolarity of fixative; (3) lymphoblasts were shown to contain antibody. Generalized cytoplasmic staining was observed in the lymphoblasts, with no localization in the ergastoplasm and with no staining of control preparations. The significance of ribosomal and lymphoblast staining is discussed in relation to cell differentiation and the development of the immune response.

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:

B-lymphocyte-induced maturation protein1 up-regulates the expression of B-cell maturation antigen in mouse plasma cells

2010 ◽  
Vol 37 (8) ◽  
pp. 3747-3755 ◽  
Author(s):  
Shaoli Deng ◽  
Tao Yuan ◽  
Xiaoxing Cheng ◽  
Rui Jian ◽  
Jing Jiang
Keyword(s):  
B Cell ◽  
B Lymphocyte ◽  
Plasma Cells ◽  
Cell Maturation ◽  
Mouse Plasma ◽  
B Cell Maturation ◽  
B Cell Maturation Antigen

Immunogold localisation of endogenous immunoglobulin-G in ultrathin frozen sections of the human placenta

1989 ◽  
Vol 257 (3) ◽  
pp. 603-607 ◽  
Author(s):  
Lopa Leach ◽  
Bryan M. Eaton ◽  
J. Anthony Firth ◽  
Soli F. Contractor
Keyword(s):  
Human Placenta ◽  
Immunoglobulin G ◽  
Frozen Sections ◽  
Immunogold Localisation ◽  
Ultrathin Frozen Sections

scholarly journals Subcellular localization of sterol carrier protein-2 in rat hepatocytes: its primary localization to peroxisomes.

1989 ◽  
Vol 108 (4) ◽  
pp. 1353-1361 ◽  
Author(s):  
G A Keller ◽  
T J Scallen ◽  
D Clarke ◽  
P A Maher ◽  
S K Krisans ◽  
...  
Keyword(s):  
Subcellular Localization ◽  
Rat Liver ◽  
Polyclonal Antibodies ◽  
Rat Hepatocytes ◽  
Carrier Protein ◽  
Sterol Carrier Protein ◽  
Rat Liver Cells ◽  
Ultrathin Frozen Sections ◽  
Sterol Carrier Protein 2

Sterol carrier protein-2 (SCP-2) is a nonenzymatic protein of 13.5 kD which has been shown in in vitro experiments to be required for several stages in cholesterol utilization and biosynthesis. The subcellular localization of SCP-2 has not been definitively established. Using affinity-purified rabbit polyclonal antibodies against electrophoretically pure SCP-2 from rat liver, we demonstrate by immunoelectron microscopic labeling of ultrathin frozen sections of rat liver that the largest concentration of SCP-2 is inside peroxisomes. In addition the immunolabeling indicates that there are significant concentrations of SCP-2 inside mitochondria, and associated with the endoplasmic reticulum and the cytosol, but not inside the Golgi apparatus, lysosomes, or the nucleus. These results were confirmed by immunoblotting experiments with proteins from purified subcellular fractions of the rat liver cells carried out with the anti-SCP-2 antibodies. The large concentration of SCP-2 inside peroxisomes strongly supports the proposal that peroxisomes are critical sites of cholesterol utilization and biosynthesis. The presence of SCP-2 inside peroxisomes and mitochondria raises questions about the mechanisms involved in the differential targeting of SCP-2 to these organelles.

Ultrathin frozen sections of yeast cells

1972 ◽  
Vol 40 (1-2) ◽  
pp. 197-204 ◽  
Author(s):  
Heinz Bauer ◽  
Elsje Sigarlakie
Keyword(s):  
Yeast Cells ◽  
Frozen Sections ◽  
Ultrathin Frozen Sections
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