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  • 1
    Electronic Resource
    Electronic Resource
    The influence of fixation on immunoperoxidase staining of plasmacells in paraffin sections of intestinal biopsy specimens (1977)
    Springer
    Histochemistry and cell biology 53 (1977), S. 57-62 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The influence of different fixation methods on the results of immunoperoxydase staining of immunoglobulin and gastrin producing cells in gastric and duodenal mucosa was investigated. An indirect method was used on paraffin sections. It appeared that that fixatives containing sublimate gave the most consistent results, a sublimate-formaldehyde mixture being the best.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00511210
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  • 2
    Electronic Resource
    Electronic Resource
    Immunoelectronmicroscopy of metaphase chromosomes (1982)
    Springer
    Histochemistry and cell biology 74 (1982), S. 341-346 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A method for the preparation of ultrathin sections of metaphase chromosomes is described. This method was applied to human metaphase chromosomes, which were immunocytochemically stained with anti-DNA and anti-ribonucleoprotein antibodies, derived from patients with auto-immune disease. Conventionally prepared metaphase spreads as well as cytocentrifuge preparations of chromosome suspensions were studied. The results indicate that the ultrastructure of chromosomes and the immunoreactivity of chromosomal constituents are influenced by the applied preparation methods. In comparison with whole mount preparations, ultrathin sections of immunostained chromosomes allow higher resolution and more precise localization of immunoreactive sites within the chromosomal structure.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00493433
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  • 3
    Electronic Resource
    Electronic Resource
    Efficiency and sensitivity of indirect immunoperoxidase methods (1983)
    Springer
    Histochemistry and cell biology 77 (1983), S. 185-194 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The peroxidase-antiperoxidase (PAP) complex method has repeatedly been claimed to be more sensitive and antibody efficient than the indirect peroxidase labeled antibody method. However, most studies comparing these methods used tissue sections as the test material. However, test systems with known amounts of antigen will allow more reliable comparison of these methods and quantitative evaluation of method sensitivity. We therefore compared the antibody efficiency and sensitivity of these methods for the detection of human chorionic gonadotropin in an enzyme linked immunosorbent assay (ELISA), an antigen spot test (AST) and tissue sections of choriocarcinoma. In the PAP technique rabbit PAP and goat anti-rabbit antibody were applied. The same antibody was peroxidase-labeled with the periodate technique and used in the labeled antibody method. In the ELISA the PAP method resulted in slightly higher antibody efficiency than the labeled antibody method. At low primary antibody dilutions the intensity of the reaction decreased with the PAP method but remained high with the labeled antibody method, in the ELISA as well as on tissue sections. In the AST the labeled antibody method and the PAP method appeared to be equally sensitive.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00506561
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  • 4
    Electronic Resource
    Electronic Resource
    FMRF-amide immunoreactivity in the mammalian gastroenteropancreatic neuroendocrine system (1986)
    Springer
    Histochemistry and cell biology 84 (1986), S. 439-444 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of FMRF-amide, a cardioactiv tetrapeptide, was studied by immunocytochemistry in human and rat gastric antrum and pancreas, and in the ovine, bovine, canine and rabbit pancreas. In human and rat gastric antrum, numerous cells contained FMRF-amide immunoreactive material. By staining of serial sections and by double staining, colocalization of immunoreactivity for gastrin and FMRF-amide was observed in part of the gastrin cells. In the pancreas of these and the other species, immunoreactivity for FMRF-amide was located both in acinar and islet endocrine cells. Colocalization of FMRF-amide and pancreatic polypeptide was found in a proportion of pancreatic polypeptide cells in the pancreas. FMRF-amide immunoreactivity never colocalized with the other neurohormonal peptides which occur in the gastric antrum and the pancreas. Our observations show that neuroendocrine cells occur in the gastric antrum and pancreas which are exclusively immunoreactive or gastrin and for pancreatic polypeptide respectively. In addition cells occur which show immunoreactivity for FMRF-amide as well as for gastrin in the gastric antrum and with antiserum to FMRF-amide as well as for pancreatic polypeptide in the pancreas. It is concluded that FMRF-amide antibodies probably recognize a substance in G and PP cells which is not identical but may be structurally related to gastrin and pancreatic polypeptide.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00482976
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  • 5
    Electronic Resource
    Electronic Resource
    Human specific anti-type IV collagen monoclonal antibodies, characterization and immunohistochemical application (1987)
    Springer
    Histochemistry and cell biology 87 (1987), S. 123-128 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This paper describes two new monoclonal antibodies reactive with human specific type IV collagen epitopes in frozen as well as routinely fixed and processed tissue sections. The antibodies (1042 and 1043) were raised against human placental type IV collagen and were shown by immunoblotting and ELISA tests to react exclusively with type IV collagen determinants. Extensive immunohistochemical survey studies on panels of tissues from various species, using unfixed cryosat sections, demonstrated that antibody 1042 reacted only with human type IV collagen whereas antibody 1043 in addition reacted with rabbit type IV collagen. All tissues showed homogeneous staining of the basement membrane, indicating that the detected epitopes did not show organ-specific distribution. Tissue processing protocols for using these monoclonal antibodies on routinely processed paraffin embedded tissues were developed. It was found that whereas polyclonal antitype IV collagen antisera required pepsin digestion, our monoclonal antibodies required pronase or papain digestion to restore type IV collagen immunoreactivity in paraffin sections. It is concluded that these monoclonal anti-type IV collagen antibodies detect species specific epitopes which can be detected in routinely processed paraffin embedded tissues after appropriate enzyme pretreatment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00533396
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  • 6
    Electronic Resource
    Electronic Resource
    Monoclonal antibodies to native basement membranes reveal heterogeneous immunoreactivity patterns (1989)
    Springer
    Histochemistry and cell biology 92 (1989), S. 407-412 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In this paper we describe the development of basement membrane (BM) reactive monoclonal antibodies (MA), by immunization of mice with intact denuded BM. The MA raised against denuded amniotic BM (clones 1052, 1053 and 1065) showed heterogeneous staining patterns. MA 1052 and 1053 reacted with epithelial BM of the epidermis and epidermal adnexa and furthermore with the epithelial alveolar BM in the lung and the superficial part of the epithelial BM in the gastrointestinal tract. MA 1065 showed immunoreactivity with the epithelial BM of epidermis and epidermal adnexa and the epithelial BM of trachea and oesophagus, and furthermore pericellular staining of the basal keratinocytes and basal corneal epithelial cells. MA 1087, raised against human glomerular BM, showed numunoreactivity with all BM, except the central epithelial BM in the cornea. The precise localization of the target epitopes in the BM was investigated on chemically cleaved human skin. Reactivity for the MA occurred predominantly in the BM lamina adherent to the dermis, suggesting that the target epitopes reside in the lamina densa and/or lamina fibroreticularis. We furthermore examined the nature of the epitopes by preincubation of tissue sections with various enzymes prior to immunohistochemistry. The reactivity of the target epitopes was not affected by bacterial collagenase, but after various protease treatments the reactivity disappeared, suggesting that the epitopes are not localized on the triple helical part of collagenous proteins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00492498
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  • 7
    Electronic Resource
    Electronic Resource
    Proliferative activity of gastric and duodenal endocrine cells in the rat (1989)
    Springer
    Histochemistry and cell biology 92 (1989), S. 325-329 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The replicative activity and migration of gastrin, somatostatin and serotonin cells in rat stomach and doudenum was studied using combined immunocytochemistry and autoradiography after 3H thymidine pulse-labeling. Our results show that a small proportion of gastrin, somatostatin and serotonin immunoreactive cells displays proliferative activity. The overall labeling index ranged from 1.3% for gastric endocrine cells to 3.2% for duodenal endocrine cells. In a pulse chase experiment, labeling indices of immunoreactive cells were estimated at several time intervals after 3H thymidine administration. Significant differences in labeling index were not found. Migration of 3H thymidine labeled endocrine cells towards the luminal surface was not found in the stomach nor in the doudenum. It is concluded that 1) these endocrine cells have replicating activity; 2) the replicative activity of endocrine cells is higher in the duodenum than in the stomach; 3) the various cell types do not show significant differences in replicating activity and 4) endocrine cells did not seem to migrate to the luminal surface of the mucosa along with the other epithelial cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00500548
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  • 8
    Electronic Resource
    Electronic Resource
    Colonic epithelium reactive monoclonal antibodies (1989)
    Springer
    Histochemistry and cell biology 92 (1989), S. 397-406 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have produced a small library of colonic mucosa and colorectal carcinoma reactive monoclonal antibodies (MoAbs) by immunizations with extracts of human colon cancer tissue and a human colon cancer cell line. Hybridoma supernatants were tested on (normal and neoplastic) human tissues by immunoperoxidase methods to evaluate organ or tissue specificity. Initial biochemical characterization of the target antigens was performed by gelpermeation chromatography, Western blotting and competition assays. Based upon the immunoreactivity patterns and the characteristics of the antigen four groups of MoAbs could be distinguished. The first group concerns the antibodies PAR-LAM 3, 9 and 10. These antibodies react with an 87 kDa protein moiety in high molecular weight (2–5×106 Da) glycoproteins. In intestinal and colon mucosa these antibodies showed diffuse binding with goblet cells. In colon carcinoma decreased reactivity with these MoAbs was found. The second group consists of antibodies PARLAM 8, 12 and 13. These antibodies react with large (〉5×106 Da) glycoproteins, most likely with carbohydrate epitopes. By immunohistochemistry in normal colon mucosa the antibodies all show granular supranuclear reactivity with goblet cells. These antibodies show increased reactivity with colon adenomas and adenocarcinomas. A third group is formed by PARLAM 2, which also reacts with a large (〉5×106 Da) glycoprotein, showing a granular distribution in goblet cells. In colon carcinomas more extensive expression is found than in normal colonic mucosa. Finally, the fourth group consists of PARLAM 11, which also reacts with a large (〉5×106 Da) glycoprotein, located in the brush border of colonic columnar cells. These antibodies might be useful tools for the analysis of the expression of mucin related glycoproteins in normal, preneoplastic and neoplastic colon mucosa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00492497
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  • 9
    Electronic Resource
    Electronic Resource
    Bromodeoxyuridine (BrdU) immunocytochemistry by exonuclease III (Exo III) digestion (1992)
    Springer
    Histochemistry and cell biology 98 (1992), S. 199-205 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A new procedure is described to generate single-stranded DNA by exonuclease III (Exo III) digestion for bromodeoxyuridine (BrdU) immunocytochemistry on tissue sections. We compared this procedure with the most widely used procedure of DNA denaturation with 2 N HCl. In vivo and in vitro pulse and continuous labelling of tissues and cells were used. The specimens were fixed in formalin, ethanol, glutaraldehyde, Carnoy's, Bouin's or Zamboni's fixative and embedded in paraffin or used unfixed as cryostat sections or cytospin preparations. After Exo III digestion, BrdU substituted DNA was detected irrespective of the fixation procedure applied. The optimal protocol for nuclease digestion appeared to be simultaneous incubation, of 10 Units Exo III per ml EcoRI buffer and anti-BrdU monoclonal antibody at 37° C. The advantages of Exo III digestion for BrdU immunocytochemistry compared to acid denaturation were: less non-specific nuclear background reactivity, no DNA renaturation, less DNA loss, optimal nuclear morphology, increase in antibody efficiency and the possibility for simultaneous detection of acid-sensitive tissue constituents. Disadvantages of the Exo III digestion are decreased sensitivity and the need for more rigorous pepsin pretreatment. We conclude that Exo III digestion of DNA is an appropriate alternative for acid denaturation for BrdU immunocytochemistry on sections of pulse-labelled specimens.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00315878
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  • 10
    Electronic Resource
    Electronic Resource
    Does ex vivo labelling of proliferating cells in colonic and vaginal mucosa reflect the S-phase fraction in vivo? (1992)
    Springer
    Histochemistry and cell biology 98 (1992), S. 267-270 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ex vivo labelling of DNA-synthesizing epithelial cells in colonic and vaginal mucosa was compared with in vivo labelling. For this purpose, in vivo S-phase cells were labelled with [3H]thymidine (Tdr) and ex vivo labelling was continued by culturing tissue specimens in bromodeoxyuridine (BrdU). Various methods of tissue culture were employed in order to improve diffusion of medium (and BrdU) in the tissue. BrdU and 3H-TdR labelling were evaluated by immunohistochemistry and autoradiography respectively. Ex vivo labelling resulted in a patchy distribution of labelled cells, which did not correspond with the 3H-TdR labelling pattern obtained in vivo. Under the described conditions ex vivo labelling does not appear to be a reliable for estimation of the proliferative activities in vivo.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00271041
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