scholarly journals ELECTRON MICROSCOPIC STUDIES OF EXPERIMENTAL NEPHRITIS WITH FERRITIN-CONJUGATED ANTIBODY

1962 ◽  
Vol 115 (5) ◽  
pp. 929-936 ◽  
Author(s):  
Giuseppe A. Andres ◽  
Councilman Morgan ◽  
Konrad C. Hsu ◽  
Richard A. Rifkind ◽  
Beatrice C. Seegal
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Rat Kidney ◽  
Fluorescent Antibody ◽  
Ultrastructural Level ◽  
Fluorescent Antibody Technique ◽  
Electron Microscopic ◽  
Antibody Technique ◽  
Experimental Nephritis ◽  
Microscopic Studies

Ferritin-conjugated antibody has been used to identify by electron microscopy the sites at which nephrotoxic globulins localize in rat kidney during acute experimental glomerulonephritis. Antibody was concentrated in the glomerular basement membrane and in basement membrane-like material contained in distended cisternae of the endoplasmic reticulum. These data confirm and amplify, at the ultrastructural level, the results of studies obtained with the fluorescent antibody technique, and are consistent with the hypothesis that the cisternae and capillary basement membrane possess common proteins.

scholarly journals ELECTRON MICROSCOPIC LOCALIZATION OF THE NEPHROTOXIC ANTIBODY IN THE GLOMERULI OF THE RAT AFTER INTRAVENOUS APPLICATION OF PURIFIED NEPHRITOGENIC ANTIBODY-FERRITIN CONJUGATES

1968 ◽  
Vol 127 (5) ◽  
pp. 867-878 ◽  
Author(s):  
Arnold Vogt ◽  
Hermann Bockhorn ◽  
Keniti Kozima ◽  
Masamichi Sasaki
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Intravenous Injection ◽  
Fluorescent Antibody ◽  
Fluorescent Antibody Technique ◽  
Electron Microscopic ◽  
Antibody Technique ◽  
Disappearance Time ◽  
Filtration Surface ◽  
Electron Microscopic Localization

Nephritis in rats was induced by intravenous injection of purified ferritin-conjugated rabbit and duck nephrotoxic globulin. Using the fluorescent antibody technique, the same capillary pattern was found as that in glomeruli of rats receiving uncoupled nephrotoxic globulin. Electron microscopy revealed a heavy accumulation of the basement membrane-fixed antibody almost exclusively at the endothelial side. A higher concentration of ferritin was demonstrable in the peripheral basement membrane. The once-fixed antibody remained at the site of reaction though decreasing with time. The half-disappearance time seemed to be shorter than that of the uncoupled nephrotoxic globulin. No difference in localization was observed between rabbit and duck antibody. At least 40 basement membrane-fixed antibody molecules from the rabbit per 3000 mµ2 of filtration surface were needed to cause immediate nephritis. To induce nephritis using duck antibody, a greater number of basement membrane-fixed antibody seemed to be necessary. No evidence of specific reaction with constituents of glomerular cells was obtained.

scholarly journals ENZYME-LABELED ANTIBODIES FOR THE LIGHT AND ELECTRON MICROSCOPIC LOCALIZATION OF TISSUE ANTIGENS

1967 ◽  
Vol 33 (2) ◽  
pp. 307-318 ◽  
Author(s):  
Paul K. Nakane ◽  
G. Barry Pierce
Keyword(s):  
Acid Phosphatase ◽  
Basement Membrane ◽  
Fluorescent Antibody ◽  
Enzymatic Reaction ◽  
Immunohistochemical Localization ◽  
Fluorescent Antibody Technique ◽  
Reaction Products ◽  
Electron Microscopic ◽  
Antibody Technique ◽  
Tissue Antigens

Enzymes, either acid phosphatase or horseradish peroxidase, were conjugated to antibodies with bifunctional reagents. The conjugates, enzymatically and immunologically active, were employed in the immunohistochemical localization of tissue antigens utilizing the reaction product of the enzymatic reaction as the marker. Tissues reacted with acid phosphatase-labeled antibodies directed against basement membrane were stained for the enzyme with Gomori's method, and those reacted with peroxidase-labeled antibody were stained with Karnovsky's method. The reaction products of the enzymes localized in the basement membrane. Unlike the preparations of the fluorescent antibody technique, enzyme-labeled antibody preparations were permanent, could be observed with an ordinary microscope, and could be examined with the electron microscope. In the latter, specific localization of antibody occurred in the basement membrane and in the endoplasmic reticulum of cells known to synthesize basement membrane antigens. The method is sensitive because of the amplifying effect of the enzymatic activity. The ultrastructural preservation and localization were better with acid phosphatase-labeled antibody than with peroxidase-labeled antibody, but acid phosphatase conjugated antibody was unstable and difficult to prepare. Peroxidase-antibody conjugates were stable and could be stored for several months at 4°C, or indefiniely in a frozen state.

Aerosol Q Fever Infection of the Nude Mouse

1981 ◽  
Vol 18 (5) ◽  
pp. 672-683 ◽  
Author(s):  
W. C. Hall ◽  
J. D. White ◽  
R. A. Kishimoto ◽  
R. E. Whitmire
Keyword(s):  
Electron Microscopy ◽  
Nude Mice ◽  
Q Fever ◽  
Fluorescent Antibody ◽  
Fluorescent Antibody Technique ◽  
Indirect Fluorescent Antibody ◽  
Antibody Technique ◽  
Athymic Nude Mice ◽  
Antibody Titers ◽  
Morphologic Changes

Athymic nude mice and euthymic littermate controls were exposed to 104 Coxiella burnetii organisms by small-particle aerosol. Antibody response with and without 2-mercaptoethanol treatment of serum was determined at various intervals after infection and serial kills were done to determine morphologic changes in both mouse phenotypes. Total antibody titers determined by the indirect fluorescent antibody technique to phase I and phase II C. burnetii were identical for both groups of mice. Microagglutinin titers determined on days 28 and 33 were abolished by 2-mercaptoethanol treatment of serum from both phenotypes, indicating that the antibody probably resided in the IgM fraction. Microscopically, the reaction to C. burnetii infection was similar in nude and euthymic mice on days 7 and 14. Later, the number and size of lesions attributable to Q fever diminished in euthymic mice. Infection was progressive in nude mice, with macrophage infiltration of most tissues, especially spleen and liver. Numerous rickettsiae were seen by immunofluorescence and electron microscopy in phagocytic vesicles of macrophages, many of which were dilated, giving the macrophage a vacuolated appearance. Results suggest that clearance of C. burnetii infection in mice is dependent upon thymus-derived lymphocytes.

scholarly journals ELECTRON MICROSCOPIC STUDIES OF EXPERIMENTAL NEPHRITIS WITH FERRITIN-CONJUGATED ANTIBODY

1963 ◽  
Vol 117 (4) ◽  
pp. 691-704 ◽  
Author(s):  
Giuseppe A. Andres ◽  
Beatrice C. Seegal ◽  
Konrad C. Hsu ◽  
Mildred S. Rothenberg ◽  
Madeleine L. Chapeau
Keyword(s):  
Bovine Serum Albumin ◽  
Basement Membrane ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Chronic Glomerulonephritis ◽  
Electron Microscopic ◽  
Antibody Technique ◽  
Microscopic Studies ◽  
Endogenous Proteins ◽  
Antigen Antibody

Acute, subacute, and chronic glomerulonephritis, similar in certain features to human glomerulonephritis, has been produced in rabbits by repeated injections of bovine serum albumin. The ratio of antigen to antibody was the factor determining the development and type of glomerulonephritis. This is in confirmation of the observations of Dixon, Feldman, and Vazquez. With the aid of the ferritin antibody technique it was shown that antigen aggregates (probably antigen-antibody complexes) are present in the blood, cross the endothelium and the basement membrane, and accumulate as dense deposits between the basement membrane and the epithelial cytoplasm. In the deposits electron-dense aggregates formed by antigen or by antigen-antibody complexes and material which might be other endogenous proteins may be identified. In rabbits dead of anaphylactic shock following injection of bovine serum albumin, dense material was found within glomerular capillaries, presumably formed by the embolic deposition of antigen-antibody complexes, since the immunofluorescein and immunoferritin techniques demonstrated the presence of both BSA and rabbit globulin.

Immunocytochemistry. A Review of Methodology for Electron Microscopic Applicaiton

1974 ◽  
Vol 32 ◽  
pp. 328-329
Author(s):  
Joseph E. Mazurkiewicz
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Ultrastructural Level ◽  
Electron Microscopic ◽  
Biological Interest ◽  
Investigative Approach ◽  
Immunologic Activity ◽  
Dense Label

Immunocytochemistry is a powerful investigative approach in which one of the most exacting examples of specificity, that of the reaction of an antibody with its antigen, isused to localize tissue and cell specific molecules in situ. Following the introduction of fluorescent labeled antibodies in T950, a large number of molecules of biological interest had been studied with light microscopy, especially antigens involved in the pathogenesis of some diseases. However, with advances in electron microscopy, newer methods were needed which could reveal these reactions at the ultrastructural level. An electron dense label that could be coupled to an antibody without the loss of immunologic activity was desired.

The blister of porphyria cutanea tarda: A Scanning and Transmission Electron Microscopic study

1986 ◽  
Vol 44 ◽  
pp. 334-335
Author(s):  
Veronika Burmeister ◽  
R. Swaminathan
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Basement Membrane ◽  
Middle Age ◽  
Porphyria Cutanea Tarda ◽  
Minor Trauma ◽  
Electron Microscopic ◽  
Transmission Electron Microscopic Study ◽  
Transmission Electron ◽  
Transmission Electron Microscopic

Porphyria cutanea tarda (PCT) is a disorder of porphyrin metabolism which occurs most often during middle age. The disease is characterized by excessive production of uroporphyrin which causes photosensitivity and skin eruptions on hands and arms, due to minor trauma and exposure to sunlight. The pathology of the blister is well known, being subepidermal with epidermodermal separation, it is not always absolutely clear, whether the basal lamina is attached to the epidermis or the dermis. The purpose of our investigation was to study the attachment of the basement membrane in the blister by comparing scanning with transmission electron microscopy.

Rapid quantification and in situ detection of nitrifying bacteria in biofilms by monoclonal antibody method

2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antibody Method ◽  
In Situ Detection ◽  
Rapid Quantification

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.

Rubella Antibodies in Human Serum: Detection by the Indirect Fluorescent-Antibody Technique

Science ◽  
1964 ◽  
Vol 145 (3635) ◽  
pp. 943-945 ◽  
Author(s):  
G. C. Brown ◽  
H. F. Maassab ◽  
J. A. Veronelli ◽  
T. J. Francis
Keyword(s):  
Human Serum ◽  
Fluorescent Antibody ◽  
Fluorescent Antibody Technique ◽  
Indirect Fluorescent Antibody ◽  
Antibody Technique ◽  
Serum Detection

scholarly journals LOCALIZATION OF UROMUCOID IN HUMAN KIDNEY AND IN SECTIONS OF HUMAN KIDNEY STONE WITH THE FLUORESCENT ANTIBODY TECHNIQUE

1965 ◽  
Vol 13 (3) ◽  
pp. 155-160 ◽  
Author(s):  
H. J. KEUTEL
Keyword(s):  
Kidney Stones ◽  
Kidney Stone ◽  
Fluorescent Antibody ◽  
Human Kidney ◽  
Fluorescent Antibody Technique ◽  
Renal Tubules ◽  
Antibody Technique ◽  
The Matrix ◽  
Normal Human ◽  
The Common

Fluorescent labeled antibodies were used for the demonstration of uromucoid. This urine specific mucoprotein is demonstrably present only in the epithelial cells of the proximal segments of the normal human renal tubules and in the matrix of human kidney stones of all the common crystalline compositions.

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