Simultaneous Immunohistochemical Demonstration of Antigen Expression and 5-bromodeoxyuridine Incorporation in Plastic Embedded Sections

Overview

Journal Histochemistry

Specialty Biochemistry

Date 1987 Jan 1

PMID 2437084

Citations 5

Authors

G Harms

H van Goor

J Koudstaal

L de Ley

M J HARDONK

Affiliations

Soon will be listed here.

Abstract

In this study a double immunohistochemical staining procedure is described for the simultaneous demonstration of antigen expressing cells and replicating cells in rat thymus. As markers for cell surface antigen expression a monoclonal antibody against Ia-expressing cells (His 19) and a monoclonal antibody against cells of the monocyte-macrophage lineage (ED2) were used. Replicating cells were demonstrated by the incorporation of 5-bromodeoxyuridine (BrdUrd). Tissue pieces were fixed in a periodate-lysine-paraformaldehyde fixative and embedded in glycol methacrylate. To demonstrate Ia-expressing cells or ED2-positive macrophages in plastic embedded sections a digestion with trypsin is necessary. The staining procedure was applied sequentially and was performed with a peroxidase and an alkaline phosphatase labeled reagent yielding respectively a brown and a blue reaction product. Results with this staining procedure on plastic embedded sections of rat thymus, an organ with a high DNA synthesizing capacity, showed incorporation of BrdUrd predominantly in the cortex. ED2-positive macrophages were only found in the cortex. The Ia-positive epithelial reticular cells demonstrated extremely well their stellate form.

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References

Beckstead J . Optimal antigen localization in human tissues using aldehyde-fixed plastic-embedded sections. J Histochem Cytochem. 1985; 33(9):954-8. DOI: 10.1177/33.9.4020104. View

Sugihara H, Hattori T, Fukuda M . Immunohistochemical detection of bromodeoxyuridine in formalin-fixed tissues. Histochemistry. 1986; 85(3):193-5. DOI: 10.1007/BF00494803. View

Sasaki K, Ogino T, Takahashi M . Immunological determination of labeling index on human tumor tissue sections using monoclonal anti-BrdUrd antibody. Stain Technol. 1986; 61(3):155-61. DOI: 10.3109/10520298609110726. View

Musiani M, Zerbini M, Plazzi M, La Placa M . Double immunoenzymatic staining for the simultaneous detection of Epstein-Barr virus induced antigens. Histochemistry. 1986; 84(1):15-7. DOI: 10.1007/BF00493414. View

Nakane P . Simultaneous localization of multiple tissue antigens using the peroxidase-labeled antibody method: a study on pituitary glands of the rat. J Histochem Cytochem. 1968; 16(9):557-60. DOI: 10.1177/16.9.557. View

Mason D, Sammons R . Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. J Clin Pathol. 1978; 31(5):454-60. PMC: 1145303. DOI: 10.1136/jcp.31.5.454. View

Takamiya H, Batsford S, Vogt A . An approach to postembedding staining of protein (immunoglobulin) antigen embedded in plastic: prerequisites and limitations. J Histochem Cytochem. 1980; 28(10):1041-9. DOI: 10.1177/28.10.6158534. View

Dijkstra C, Dopp E, Joling P, Kraal G . The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3. Immunology. 1985; 54(3):589-99. PMC: 1453512. View

Harms G, van Goor H, Koudstaal J, de Ley L, HARDONK M . Immunohistochemical demonstration of DNA-incorporated 5-bromodeoxyuridine in frozen and plastic embedded sections. Histochemistry. 1986; 85(2):139-43. DOI: 10.1007/BF00491761. View

10.

Erlandsen S, Hegre O, Parsons J, McEvoy R, Elde R . Pancreatic islet cell hormones distribution of cell types in the islet and evidence for the presence of somatostatin and gastrin within the D cell. J Histochem Cytochem. 1976; 24(7):883-97. DOI: 10.1177/24.7.60437. View

11.

Huang S, Minassian H, More J . Application of immunofluorescent staining on paraffin sections improved by trypsin digestion. Lab Invest. 1976; 35(4):383-90. View

12.

Campbell G, Bhatnagar A . Simultaneous visualization by light microscopy of two pituitary hormones in a single tissue section using a combination of indirect immunohistochemical methods. J Histochem Cytochem. 1976; 24(2):448-52. DOI: 10.1177/24.2.1254941. View

13.

STERNBERGER L, Joseph S . The unlabeled antibody method. Contrasting color staining of paired pituitary hormones without antibody removal. J Histochem Cytochem. 1979; 27(11):1424-9. DOI: 10.1177/27.11.92498. View

14.

Falini B, De Solas I, Halverson C, Parker J, Taylor C . Double labeled-antigen method for demonstration of intracellular antigens in paraffin-embedded tissues. J Histochem Cytochem. 1982; 30(1):21-6. DOI: 10.1177/30.1.6798106. View

15.

McLean I, Nakane P . Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974; 22(12):1077-83. DOI: 10.1177/22.12.1077. View

16.

Martin-Comin J, Robyn C . Comparative immunoenzymatic localization of prolactin and growth hormone in human and rat pituitaries. J Histochem Cytochem. 1976; 24(9):1012-6. DOI: 10.1177/24.9.61240. View