scholarly journals Application of the FITC-anti-FITC-gold system to ultrastructural localization of antigens.

1991 ◽  
Vol 39 (12) ◽  
pp. 1725-1728 ◽  
Author(s):  
G J van Dam ◽  
B J Bogitsh ◽  
J A Fransen ◽  
D Kornelis ◽  
R J van Zeyl ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Fluorescein Isothiocyanate ◽  
High Specificity ◽  
Ultrastructural Localization ◽  
Good Alternative ◽  
Ultrastructural Level ◽  
Electron Microscopic ◽  
Protein Conjugates ◽  
Specificity And Sensitivity ◽  
Microscopic Studies

We report the application of a fluorescein isothiocyanate (FITC)-anti-FITC method to localize antigens at the ultrastructural level. In the systems studied, the anti-FITC-based detection method displays high specificity and sensitivity. These observations, combined with ease of production and with availability of FITC-protein conjugates, suggest that the FITC-anti-FITC method is a good alternative to presently used methods and is widely applicable to immunochemical and immunocytochemical procedures. The same preparation and protocol can be used for light and electron microscopic studies, thereby reducing possible artifacts introduced if different procedures are used. In the present study, two systems were used to test the method. One system used an FITC-labeled monoclonal antibody (MAb) to schistosome circulating cathodic antigen. In this system, the label was detected in the gut of adult Schistosoma mansoni by an anti-FITC MAb conjugated to 10-nm gold particles. The second system used human IgM antibodies pooled from patients infected with Schistosoma mansoni. In this system detection was accomplished using an anti-human IgM-FITC conjugate followed by the anti-FITC-Au antibody conjugate.

Peroxidase Localization in Hartmanella Trophozoites

1971 ◽  
Vol 29 ◽  
pp. 346-347 ◽  
Author(s):  
George E. Childs ◽  
Joseph H. Miller
Keyword(s):  
Hydrogen Peroxide ◽  
Ultrastructural Localization ◽  
Pathogenic Strain ◽  
Oxidative Enzymes ◽  
Differential Centrifugation ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
The Subject ◽  
Axenic Cultures

Biochemical and differential centrifugation studies have demonstrated that the oxidative enzymes of Acanthamoeba sp. are localized in mitochondria and peroxisomes (microbodies). Although hartmanellid amoebae have been the subject of several electron microscopic studies, peroxisomes have not been described from these organisms or other protozoa. Cytochemical tests employing diaminobenzidine-tetra HCl (DAB) and hydrogen peroxide were used for the ultrastructural localization of peroxidases of trophozoites of Hartmanella sp. (A-l, Culbertson), a pathogenic strain grown in axenic cultures of trypticase soy broth.

scholarly journals Comparative parasitological and electron microscopic studies on the effects of Nitazoxanide and Praziquantel in Schistosoma mansoni-infected mice

2016 ◽  
Vol 4 (2) ◽  
pp. 228
Author(s):  
Hend A. El-Taweel ◽  
Mona H. El-Sayad ◽  
Sahar A. Abu Helw ◽  
Mohammad A. Al-Kazzaz
Keyword(s):  
Schistosoma Mansoni ◽  
Control Group ◽  
Electron Microscopic ◽  
Male And Female ◽  
Scanning Electron Microscopic ◽  
Antischistosomal Activity ◽  
Microscopic Studies ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

This study was designed to evaluate antischistosomal activity of Nitazoxanide (NTZ) in Schistosoma mansoni-infected mice compared to Praziquantel (PZQ). Fifty four infected mice were recruited into 3 groups, each of 18 mice. Group 1 was infected non-treated control. Group 2 was infected and then treated with PZQ 500 mg for two days, and group 3 was infected and treated with NTZ 100 mg/kg for seven days. Efficacy of drugs was assessed by Parasitological, and scanning electron microscopic studies. PZQ reduced (4.9%, 22.5% and 50.7%) of faecal eggs, (22%, 22.6% and 55.1%) of intestinal eggs, (20.4%, 44.3% and 46.7%) of hepatic egg counts and (27%, 45.1% and 64.9%) of total worm load whereas, NTZ reduced (4.9%, 22.5% and 50.7%),of faecal eggs, (22%, 22.6% and 55.1%) of intestinal eggs ,(20.4%, 44.3% and 46.7%) of hepatic egg counts and (27%, 45.1% and 64.9%) of total worm load at 1, 2 and 4 WPT, respectively. The percentages of dead eggs were more than 80% after PZQ treatment and only 30% after NTZ at 4 WPT. PZQ showed extensive tegumental damages in male and female worms more than NTZ at 2 WPT. Our findings concluded that Nitazoxanide showed weaker antischistosomal activity in animal models than praziquantel.

scholarly journals Host cell adhesion to Schistosoma mansoni larvae in the peritoneal cavity of naive mice: histological and scanning electron microscopic studies

1993 ◽  
Vol 35 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Alan Lane de Melo ◽  
Conceição Ribeiro da Silva Machado ◽  
Leógenes Horácio Pereira
Keyword(s):  
Cell Adhesion ◽  
Schistosoma Mansoni ◽  
Host Cell ◽  
Peritoneal Cavity ◽  
Mononuclear Cells ◽  
Close Contact ◽  
Histological Study ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
Scanning Electron

Cercariae of Schistosoma mansoni inoculated into the peritoneal cavity of naive mice induced host cell adhesion to their surface, but after 90 minutes the number of adherent cells sharply decreased. The cell detachment is progressive and simultaneous to the cercaria-schistosomule transformation. The histological study showed mainly neutrophils in close contact with the larvae. Mononuclear cells and some eosinophils were occasionally seen surrounding the adherent neutrophils. The scanning electron microscopy showed cells displaying twisted microvilli and several microplicae contacting or spreading over the larval surface, and larvae completely surrounded by clusters of cells. These results suggest that the neutrophils recognize molecules on the cercarial surface which induce their spreading

scholarly journals Visualization of anti-sperm plasma membrane IgG and Fab as a method for localization of boar sperm membrane antigens.

1982 ◽  
Vol 30 (12) ◽  
pp. 1217-1227 ◽  
Author(s):  
L D Russell ◽  
R N Peterson ◽  
T A Russell
Keyword(s):  
Plasma Membrane ◽  
Protein A ◽  
Fluorescein Isothiocyanate ◽  
Ultrastructural Localization ◽  
Surface Antigens ◽  
Ultrastructural Level ◽  
Simple Method ◽  
Sperm Surface ◽  
Sperm Plasma Membrane ◽  
Sperm Antigens

A simple method for ultrastructural localization of sperm surface antigens by direct visualization of bound antibodies is presented. Anti-sperm plasma membrane (ASPM) immunoglobulin (Ig) G, visualized in tissues treated with an osmium:ferrocyanide mixture, projected 11-13 nm from the surface and ASPM Fab fragments projected 8-10 nm from the surface. The density of IgG labeling, as subjectively estimated, corresponded to indirect immune fluorescein isothiocyanate, indirect immunoferritin, and sperm-vesicle labeling patterns. Agglutination of sperm vesicles and sperm were demonstrated and the linking antibody visualized. A second antibody on protein A directed against ASPM IgG made the immunologic tag more apparent and indicated, in disrupted sperm preparations, labeling of both sides of the plasma membrane. The method provides for easy and sensitive localization of sperm surface antigens at the ultrastructural level and is presently being used to localize specific sperm antigens.

scholarly journals Correlative immunofluorescence and electron microscopy on the same section of epon-embedded material.

1985 ◽  
Vol 33 (2) ◽  
pp. 165-171 ◽  
Author(s):  
C L Rieder ◽  
S S Bowser
Keyword(s):  
Electron Microscopy ◽  
Core Protein ◽  
Fluorescein Isothiocyanate ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Immunofluorescent Localization ◽  
Cellular Inclusions ◽  
Tumor Virus ◽  
Specific Antigens ◽  
First Time

Semithick (0.25-0.50 micron) sections, cut from cells stained with fluorescein isothiocyanate (FITC)-conjugated antibodies prior to embedding in Epon, show high resolution patterns of immunofluorescence against a background void of autofluorescence. These same sections can then be viewed, after uranyl and lead staining, in the electron microscope. We clearly establish the specificity of this same-section correlative immunofluorescence-electron microscopy approach by showing that the immunofluorescent patterns observed in such sections of cells, stained prior to embedding for the indirect immunofluorescent localization of tubulin, follows the distribution of microtubules within the same sections as determined by electron microscopy. We then use this method to demonstrate for the first time that the 57 kD core protein of wound tumor virus is associated, at the ultrastructural level, with two distinct cellular inclusions in virally infected AC-20 cells. In some instances the fidelity in the correlation between the distribution of immunofluorescently labeled antigens and the ultrastructure in the same section eliminates the need to employ more complex procedures for labeling antigens for ultrastructural detection. This technique, therefore, provides a rapid and simple first approach to many problems that require the ultrastructural localization of specific antigens.

scholarly journals Analysis of opticin binding to collagen fibrils identifies a single binding site in the gap region and a high specificity towards thin heterotypic fibrils containing collagens II, and XI or V/XI

2020 ◽  
Author(s):  
Uwe Hansen ◽  
David F. Holmes ◽  
Peter Bruckner ◽  
Paul N. Bishop
Keyword(s):  
Binding Site ◽  
High Specificity ◽  
Collagen Fibrils ◽  
Class Iii ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
Collagen Ii ◽  
Fibril Morphology ◽  
And Cartilage

AbstractOpticin is a class III member of the extracellular matrix small leucine-rich repeat protein/proteoglycan (SLRP) family found in vitreous humour and cartilage. It was first identified associated with the surface of vitreous collagen fibrils and several other SLRPs are also known to bind collagen fibrils and it some cases alter fibril morphology. The purpose of this study was to investigate the binding of opticin to the collagen II-containing fibrils found in vitreous and cartilage. Electron microscopic studies using gold labelling demonstrated that opticin binds vitreous and thin cartilage collagen fibrils specifically at a single site in the gap region of the collagen D-period corresponding to the e2 stain band; this is the first demonstration of the binding site of a class III SLRP on collagen fibrils. Opticin did not bind thick cartilage collagen fibrils from cartilage or tactoids formed in vitro from collagen II, but shows high specificity for thin, heterotypic collagen fibrils containing collagens II, and XI or V/XI. Vitreous collagen fibrils from opticin null and wild-type mice were compared and no difference in fibril morphology or diameter was observed. Similarly, in vitro fibrillogenesis experiments showed that opticin did not affect fibril formation. We propose that when opticin is bound to collagen fibrils, rather than influencing their morphology it instead hinders the binding of other molecules to the fibril surfaces and/or act as an intermediary bridge linking the collagen fibrils to other non-collagenous molecules.

Ultrastructural localization of Sm28 GST protective antigen in Schistosoma mansoni adult worms

Parasitology ◽  
1996 ◽  
Vol 113 (4) ◽  
pp. 377-391 ◽  
Author(s):  
J. L. Liu ◽  
J. Fontaine ◽  
A. Capron ◽  
J. M. Grzych
Keyword(s):  
Schistosoma Mansoni ◽  
Protective Antigen ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Staining Intensity ◽  
Wide Distribution ◽  
Published Data ◽  
Glutathione S Transferase ◽  
Sustentacular Cells ◽  
Germinal Cells

SUMMARYThe localization of the 28 kDa Schistosoma mansoni glutathione S-transferase (Sm28 GST) has been investigated using immunohistochemistry and electron microscopy and the results compared with previously published data. This study confirms the wide distribution of this antigen in the parasite. In male and female worms, Sm28 GST is localized in the tegument, the parenchyma, the oesophageal epithelium and in genital organs. Sm28 GST was clearly detected in germinal and sustentacular cells. The decrease of staining intensity during the differentiation of germinal cells suggests a down-regulated expression of the molecule. At the ultrastructural level, this antigen was abundant in nuclei and less present in the cytoplasm. The marked heterogeneity observed in the staining of individual worms indicates that Sm28 GST seems to be closely associated with the parasite’s metabolism. The results are discussed in relation to the biological and protective functions of the protein.

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.

Sponge secondary metabolites: biochemical and ultrastructural localization of the antimitotic agent avarol in Dysidea avara.

1986 ◽  
Vol 34 (12) ◽  
pp. 1687-1690 ◽  
Author(s):  
W E Müller ◽  
B Diehl-Seifert ◽  
C Sobel ◽  
A Bechtold ◽  
Z Kljajić ◽  
...  
Keyword(s):  
Ultrastructural Localization ◽  
Inhibitory Effect ◽  
Storage Site ◽  
Electron Microscopic ◽  
Antimitotic Agent ◽  
Transmission Electron ◽  
Cytoplasmic Vesicles ◽  
Transmission Electron Microscopic ◽  
Microscopic Studies ◽  
Dysidea Avara

The secondary metabolite avarol, a potent cytostatic and antibacterial sesquiterpenoid hydroquinone, is present in large amounts only in the sponge Dysidea avara (2.7 g avarol/1 kg of fresh material). The present study was designed to determine the storage site of this compound within the organism. Light and transmission electron microscopic studies revealed that avarol is probably stored only in spherular cells. The compound is compartmented in intracellular cytoplasmic vesicles in a paracrystalline form, and therefore can have no inhibitory effect on the sponge cells. Quantitative analysis utilizing high-pressure liquid chromatography revealed that avarol is present at a concentration of 3.2 micrograms/10(6) spherular cells. It appears that avarol is released from the cells into the extracellular space in a merocrine manner. We suggest that it is involved in regulating the bacteria with which the sponge is symbiotically associated.

scholarly journals Correlative light and electron microscopic immunocytochemistry on the same section with colloidal gold.

1987 ◽  
Vol 35 (4) ◽  
pp. 419-425 ◽  
Author(s):  
H Mar ◽  
T Tsukada ◽  
A M Gown ◽  
T N Wight ◽  
D G Baskin
Keyword(s):  
Colloidal Gold ◽  
High Specificity ◽  
Ultrastructural Localization ◽  
Electron Microscopic ◽  
Electron Microscopic Immunocytochemistry ◽  
Antigenic Sites ◽  
Embedding Technique ◽  
Positive Immunostaining ◽  
Arterial Smooth Muscle Cells ◽  
Electron Microscopic Localization

Ultrastructural localization of growth hormone in rat anterior pituitary and of muscle-specific actin in rabbit arterial smooth muscle cells was accomplished with a post-embedment procedure using colloidal gold. Plastic sections (2 microns) were mounted on slides, deplasticized, immunostained with immunoglobulin-colloidal gold particles, re-embedded in Epon, and sectioned for electron microscopy. This procedure enabled light and electron microscopic localization of these intracellular antigens on the same section. Positive immunostaining was demonstrated with this procedure with a muscle-specific actin antibody which previously failed to localize antigenic sites by EM. The procedure described yielded staining of high specificity, with minimal background and well-preserved ultrastructure. This re-embedding technique is useful in situations where problems with post-embedding EM immunostaining exist and where correlative LM and EM immunostaining is essential.

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