scholarly journals ULTRASTRUCTURAL IMMUNOCYTOCHEMICAL LOCALIZATION OF LYSOZYME IN THE PANETH CELLS OF MAN

1974 ◽  
Vol 22 (6) ◽  
pp. 401-413 ◽  
Author(s):  
STANLEY L. ERLANDSEN ◽  
JONATHAN A. PARSONS ◽  
THOMAS D. TAYLOR
Keyword(s):  
Paneth Cell ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Paneth Cells ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Specific Staining ◽  
Secretion Granules ◽  
Normal Sheep Serum ◽  
Unlabeled Antibody

Human lysozyme was localized immunocytochemically at the ultrastructural level within Paneth cells of man by use of the unlabeled antibody enzyme method. Specific staining for lysozyme was observed over secretion granules in the apical cytoplasm, within the region of the Golgi apparatus and within some, but not all, lysosomes. No staining was observed within the cisternae of the rough endoplasmic reticulum or other cellular organelles. In control experiments comparing semiadjacent sections of the same Paneth cell, substitution of either normal rabbit serum for rabbit antihuman lysozyme antiserum (specificity control) or normal sheep serum for sheep antirabbit immunoglobulin G antiserum (method control) completely eliminated specific staining for lysozyme. The intensity of staining for lysozyme was related to both the titer and length of exposure to antilysozyme antiserum. Specific staining was obtained in tissue embedded in Araldite or Epon and was facilitated by etching with hydrogen peroxide. No staining was observed after prolonged fixation in glutaraldehyde or treatment with uranyl acetate in block.

scholarly journals Immunocytochemical identification and localization of immunoglobulin A within Paneth cells of the rat small intestine.

1976 ◽  
Vol 24 (10) ◽  
pp. 1085-1092 ◽  
Author(s):  
S L Erlandsen ◽  
C B Rodning ◽  
C Montero ◽  
J A Parsons ◽  
E A Lewis ◽  
...  
Keyword(s):  
Light Chain ◽  
Cell Function ◽  
Paneth Cell ◽  
Immunoglobulin A ◽  
Cross Reactivity ◽  
Paneth Cells ◽  
Immunoglobulin Light Chain ◽  
Specific Staining ◽  
Heavy Chains ◽  
Purified Antigen

Light microscopic immunocytochemistry was used to identify Paneth cells by their lysozyme content and to detect immunoglobulin antigens within a subpopulation of these cells. Antisera specific for the heavy chains of rat or human immunoglobulin A and for immunoglobulin light chain antigens produced specific staining of rat Paneth cells. The distribution of immunoglobulin staining varied between adjacent Paneth cells in the same crypt and between Paneth cells in adjacent crypts, as well as between Paneth cell populations of different animals. No staining of rat Paneth cells was detected using antisera specific for the heavy chain of immunoglobulins G or M. The specific staining of Paneth cells for immunoglobulin A and light chain antigens was blocked by absorption of each antiserum with its respective purified antigen. Absorption of these antisera with purified rat lysozyme did not affect staining and thereby eliminated the possibility of immunologic cross-reactivity between lysozyme and immunoglobulin antigens. It is suggested, in light of current concepts of Paneth cell function, that the immunoglobulin staining of Paneth cells may reflect their ability to phagocytize immunoglobulin A-coated microorganisms or immune complexes containing immunoglobulin A.

scholarly journals ULTRASTRUCTURAL LOCALIZATION OF A SOLUBLE COLLAGEN ANTIGEN

1971 ◽  
Vol 19 (11) ◽  
pp. 663-669 ◽  
Author(s):  
LIVIA LUSTIG
Keyword(s):  
Chick Embryo ◽  
Light And Electron Microscopy ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Collagen Fibrils ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Neutral Salt ◽  
Antibody Technique ◽  
Soluble Collagen

Soluble collagen antigen was localized at the ultrastructural level using an enzyme-labeled antibody technique. Specific antiserum against neutral salt-soluble collagen was conjugated with horseradish peroxidase. Tissue culture of fibroblasts, sections of chick embryo connective tissue and skin and tendon of newborn and adult chickens were incubated with the conjugated antiserum and processed for light and electron microscopy. Light microscopic observations showed a localization of the collagen antigen in the cytoplasm of fibroblasts and in the extracellular collagen fibers. Ultrastructurally, the antigen was found on the ribosomes, on the membranes of the rough endoplasmic reticulum, in the intracisternal material of fibroblasts and extracellularly in the collagen fibrils. The collagen fibrils of chick embryo cartilage were intensely stained while the matrix itself remained unstained. Controls performed with absorbed antiserum, conjugated normal rabbit serum or different antigens failed to react with the tissues.

Ultrastructural localization of viral antigens using the unlabeled antibody-enzyme method.

1976 ◽  
Vol 24 (3) ◽  
pp. 517-526 ◽  
Author(s):  
G Wendelschafer-Crabb ◽  
S L Erlandsen ◽  
D H Walker
Keyword(s):  
Staining Method ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Model System ◽  
Shigella Dysenteriae ◽  
Specific Staining ◽  
Bacteriophage P1 ◽  
Viral Antigens ◽  
Enzyme Method ◽  
Unlabeled Antibody

Employing the unlabeled antibody enzyme method at the ultrastructural level, a comparison was made between preembedding staining and postembedding staining for the detection of viral antigens. The bacteriophage P1 absorbed to the surface of Shigella dysenteriae was used as a model system. Preembedding staining resulted in the specific deposition of peroxidase-antiperoxidase (PAP) complexes as an electron-dense coating around the viral heads. Disadvantages of the preembedding staining method included the agglutination of cells by the primary antiserum which produced a gradient of specific staining and the "bleeding" or migration of electron dense reaction product away from the sites of attached PAP complexes. The postembedding staining method had distinct advantages over the preembedding staining in that PAP complexes were deposited directly over exposed viral heads within the thin section. In addition, the specific immunostaining of viruses was uniform through the section and no artifactual migration of reaction product was observed.

Ultrastructural changes in sciatic nerve tissue: Effects of passive maternal smoking

1983 ◽  
Vol 41 ◽  
pp. 650-651
Author(s):  
Amankwah K.S. ◽  
A.D. Weberg ◽  
R.C. Kaufmann
Keyword(s):  
Sciatic Nerve ◽  
Maternal Smoking ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Nerve Tissue ◽  
Ultrastructural Changes ◽  
Cacodylate Buffer ◽  
And Control ◽  
Spontaneous Delivery

Previous research has revealed that passive (involuntary inhalation) tobacco smoking during gestation can have adverse effects upon the developing fetus. These prior investigations did not concentrate on changes in fetal morphology. This study was undertaken to delineate fetal neural abnormalities at the ultrastructural level in mice pups exposed in utero to passive maternal smoking.Pregnant study animals, housed in a special chamber, were subjected to cigarette smoke daily from conception until delivery. Blood tests for determination of carbon monoxide levels were run at 15-18 days gestation. Sciatic nerve tissue from experimental and control animals were obtained following spontaneous delivery and fixed in 2.5% gluteraldehyde in 0.1M cacodylate buffer pH 7.3. The samples were post-fixed in osmium ferrocyanide (1:1 mixture of 1.5% aqueous OSO4 and 2.5% K4 Fe(CN)6). Following dehydration, the tissues were infiltrated with and embedded in Spurr. Sections were stained with uranyl acetate and lead citrate.

The Ultrastructure of Metastatic Human Mammary Carcinoma After Prolonged Estrogen Therapy

1967 ◽  
Vol 25 ◽  
pp. 180-181
Author(s):  
John H.L. Watson ◽  
John L. Swedo ◽  
R.W. Talley
Keyword(s):  
Mammary Carcinoma ◽  
Osmium Tetroxide ◽  
Estrogen Therapy ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Human Mammary Carcinoma ◽  
Surgical Biopsy ◽  
Lead Hydroxide ◽  
Preliminary Study ◽  
Hormonal Therapies

A preliminary study of human mammary carcinoma on the ultrastructural level is reported for a metastatic, subcutaneous nodule, obtained as a surgical biopsy. The patient's tumor had responded favorably to a series of hormonal therapies, including androgens, estrogens, progestins, and corticoids for recurring nodules over eight years. The pertinent nodule was removed from the region of the gluteal maximus, two weeks following stilbestrol therapy. It was about 1.5 cms in diameter, and was located within the dermis. Pieces from it were fixed immediately in cold fixatives: phosphate buffered osmium tetroxide, glutaraldehyde, and paraformaldehyde. Embedment in each case was in Vestopal W. Contrasting was done with combinations of uranyl acetate and lead hydroxide.

An ultrastructural study of sertoli cells in two geographically isolated populations of Aphanius dispar

1992 ◽  
Vol 50 (1) ◽  
pp. 548-549
Author(s):  
Taber A. Ba-Omar ◽  
Philip F. Prentis
Keyword(s):  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Freshwater Teleost ◽  
Isolated Populations ◽  
En Bloc ◽  
The North ◽  
Group A ◽  
Ultrathin Sections ◽  
Group B ◽  
Geographically Isolated

We have recently carried out a study of spermiogenic differentiation in two geographically isolated populations of Aphanius dispar (freshwater teleost), with a view to ascertaining variation at the ultrastructural level. The sampling areas were the Jebel Al Akhdar in the north (Group A) and the Dhofar region (Group B) in the south. Specimens from each group were collected, the testes removed, fixed in Karnovsky solution, post fixed in OsO, en bloc stained with uranyl acetate and then routinely processed to Agar 100 resin, semi and ultrathin sections were prepared for study.

Identification of Maize Rayado Fino Virus by Immunoelectron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 636-637
Author(s):  
O. E. Bradfute
Keyword(s):  
Electron Microscopy ◽  
Zea Mays ◽  
Costa Rica ◽  
Severe Disease ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Virus Particles ◽  
Far North ◽  
Maize Rayado Fino Virus ◽  
Antibody Coating

Maize rayado fino virus (MRFV) causes a severe disease of corn (Zea mays) in many locations throughout the neotropics and as far north as southern U.S. MRFV particles detected by direct electron microscopy of negatively stained sap from infected leaves are not necessarily distinguishable from many other small isometric viruses infecting plants (Fig. 1).Immunosorbent trapping of virus particles on antibody-coated grids and the antibody coating or decoration of trapped virus particles, was used to confirm the identification of MRFV. Antiserum to MRFV was supplied by R. Gamez (Centro de Investigacion en Biologia Celular y Molecular, Universidad de Costa Rica, Ciudad Universitaria, Costa Rica).Virus particles, appearing as a continuous lawn, were trapped on grids coated with MRFV antiserum (Fig. 2-4). In contrast, virus particles were infrequently found on grids not exposed to antiserum or grids coated with normal rabbit serum (similar to Fig. 1). In Fig. 3, the appearance of the virus particles (isometric morphology, 30 nm diameter, stain penetration of some particles, and morphological subunits in other particles) is characteristic of negatively stained MRFV particles. Decoration or coating of these particles with MRFV antiserum confirms their identification as MRFV (Fig. 4).

Ultrastructural Localization of Antigens by Capillary Action Immunocytochemistry

1996 ◽  
Vol 54 ◽  
pp. 40-41
Author(s):  
László G. Kömüves
Keyword(s):  
San Francisco ◽  
Colloidal Gold ◽  
Specific Binding ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Adhesive Tape ◽  
Distilled Water ◽  
Capillary Action ◽  
Rabbit Igg

Light microscopic immunohistochemistry based on the principle of capillary action staining is a widely used method to localize antigens. Capillary action immunostaining, however, has not been tested or applied to detect antigens at the ultrastructural level. The aim of this work was to establish a capillary action staining method for localization of intracellular antigens, using colloidal gold probes.Post-embedding capillary action immunocytochemistry was used to detect maternal IgG in the small intestine of newborn suckling piglets. Pieces of the jejunum of newborn piglets suckled for 12 h were fixed and embedded into LR White resin. Sections on nickel grids were secured on a capillary action glass slide (100 μm wide capillary gap, Bio-Tek Solutions, Santa Barbara CA, distributed by CMS, Houston, TX) by double sided adhesive tape. Immunolabeling was performed by applying reagents over the grids using capillary action and removing reagents by blotting on filter paper. Reagents for capillary action staining were from Biomeda (Foster City, CA). The following steps were performed: 1) wet the surface of the sections with automation buffer twice, 5 min each; 2) block non-specific binding sites with tissue conditioner, 10 min; 3) apply first antibody (affinity-purified rabbit anti-porcine IgG, Sigma Chem. Co., St. Louis, MO), diluted in probe diluent, 1 hour; 4) wash with automation buffer three times, 5 min each; 5) apply gold probe (goat anti-rabbit IgG conjugated to 10 nm colloidal gold, Zymed Laboratories, South San Francisco, CA) diluted in probe diluent, 30 min; 6) wash with automation buffer three times, 5 min each; 7) post-fix with 5% glutaraldehyde in PBS for 10 min; 8) wash with PBS twice, 5 min each; 9) contrast with 1% OSO4 in PBS for 15 min; 10) wash with PBS followed by distilled water for5 min each; 11) stain with 2% uranyl acetate for 10 min; 12) stain with lead citrate for 2 min; 13) wash with distilled water three times, 1 min each. The glass slides were separated, and the grids were air-dried, then removed from the adhesive tape. The following controls were used to ensure the specificity of labeling: i) omission of the first antibody; ii) normal rabbit IgG in lieu of first antibody; iii) rabbit anti-porcine IgG absorbed with porcine IgG.

Ultrastructural morphology of neurosecretory neurons in the barnacle Balanus amphitrite

1990 ◽  
Vol 48 (3) ◽  
pp. 434-435
Author(s):  
Grazia Tagliafierro ◽  
Cristiana Crosa ◽  
Marco Canepa ◽  
Tiziano Zanin
Keyword(s):  
Nervous System ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Balanus Amphitrite ◽  
Neurosecretory Neurons ◽  
The Central Nervous System ◽  
Ultrathin Sections ◽  
Dense Core Granules ◽  
The Brain ◽  
Ocellar Nerve

Barnacles are very specialized Crustacea, with strongly reduced head and abdomen. Their nervous system is rather simple: the brain or supra-oesophageal ganglion (SG) is a small bilobed structure and the toracic ganglia are fused into a single ventral mass, the suboesophageal ganglion (VG). Neurosecretion was shown in barnacle nervous system by histochemical methods and numerous putative hormonal substances were extracted and tested. Recently six different types of dense-core granules were visualized in the median ocellar nerve of Balanus hameri and serotonin and FMRF-amide like substances were immunocytochemically detected in the nervous system of Balanus amphitrite. The aim of the present work is to localize and characterize at ultrastructural level, neurosecretory neuron cell bodies in the VG of Balanus amphitrite.Specimens of Balanus amphitrite were collected in the port of Genova. The central nervous system were Karnovsky fixed, osmium postfixed, ethanol dehydrated and Durcupan ACM embedded. Ultrathin sections were stained with uranyl acetate and lead citrate. Ultrastructural observations were made on a Philips M 202 and Zeiss 109 T electron microscopy.

Localization of oxytocin and neurophysin in baboon (Papio anubis) corpus luteum by immunocytochemistry

1986 ◽  
Vol 113 (4) ◽  
pp. 570-575 ◽  
Author(s):  
Firyal S. Khan-Dawood
Keyword(s):  
Corpus Luteum ◽  
Rabbit Serum ◽  
Corpora Lutea ◽  
Positive Staining ◽  
Normal Rabbit Serum ◽  
Fallopian Tubes ◽  
Papio Anubis ◽  
Luteal Cells ◽  
Luteal Tissue ◽  
Immunocytochemical Reaction

Abstract. Immunoreactive oxytocin is detectable in the corpora lutea of women and cynomolgus monkeys by radioimmunoassay. To localize the presence of oxytocin and neurophysin I in ovarian tissues of subhuman primates, three corpora lutea and ovarian stromal tissues and two Fallopian tubes obtained during the menstrual cycle of the baboon and decidua from two pregnant baboons were examined using highly specific antisera against either oxytocin or neurophysin I and preoxidase-antiperoxidase light microscopy immunohistochemistry. Oxytocin-like as well as neurophysin I-like immunoreactivities were found in some cells of all the corpora lutea only, but could not be demonstrated in ovarian stromal tissues, Fallopian tubes and decidua. Specificity of the immunocytochemical reaction was further confirmed by immunoabsorption of the antiserum with excess oxytocin or neurophysin, after which the immunoreactivities for both oxytocin and neurophysin in the luteal tissue were negative. Similar controls using normal rabbit serum gave no positive staining for either oxytocin or neurophysin. Counterstaining of the positive immunoreactivities for oxytocin and neurophysin I with Mayer's haematoxylin and eosin demonstrated clearly that the oxytocin and neurophysin I appeared as granular material mainly within the cytoplasm of the luteal cells. The localization of immunoreactive oxytocin and neurophysin I in the corpus luteum of the baboon demonstrates directly the presence of these two neurohypophysial peptides within primate luteal cells and suggests their local production.

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