Summary
The usefulness of 1-naphthol as substrate for horseradish peroxidase (HRP) in immunohistochemistry was studied using the peroxidase-antiperoxidase (PAP) and avidin-biotin-complex (ABC) methods in the demonstration of glial fibrillary acidic protein (GFAP), vimentin, carbonic anhydrase C (CA.C), and factor VIII-related antigen (FVIII/RAg) in central nervous tissue and cerebral tumors. In the presence of ammonium carbonate, 1-naphthol is oxidized by HRP and hydrogen peroxide, producing a fine gray-violet precipitate. The oxidation product of 1-naphthol proved capable of binding a great number of basic dyes. For each stain the final reaction product had a characteristic color that was different from the spontaneous color of the dye and from the color displayed by nuclei. The final color obtained with this procedure was alcohol resistant and could be mounted in solvent-based mounting media. The results obtained with the 1-nappthol basic dye (1-NBD) method were compared with those obtained using the diaminobenzidine (DAB) reaction in the demonstration of GFAP-positive astrocytes. The DAB reaction produced a more intense staining but also a coarser precipitate than the 1-NBD reaction. The 1-NBD procedure showed more morphological detail of fine structures and did not obscure nuclei and mitosis. The very low toxicity of 1-naphthol compared with DAB (a suspected carcinogen) is an important advantage of the 1-NBD method, as is its high specificity and sensitivity.
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References
Brignac PJ, Patel V (1974) The oxidation of 2-naphthol, 1-naphthol and 2,7-dihydroxynaphthalene by horseradish peroxidase and hydrogen peroxide. Anal Lett 7:195–203
Burnstone MS (1962) Enzyme histochemistry and its application in the study of neoplasms. Academic Press, New York, pp 452–454
Deane HW, Barnett RJ, Selingman AM (1960) Histochemische Methoden zum Nachweis der Enzymaktivität. In: Graumann W, Neumann K (eds) Handbuch der Histochemie. Vol VII: Enzyme. G Fischer, Stuttgart, p 39
De Bruyn PPH (1939) Phenol granulation of leucocytes and its staining with specific lipoid stains (Sudan). Acta Need Morphol 2:322–344
Finar JL (1963) Organic chemistry. Vol 1: The fundamental principles. 4th edn. Longmans, London
Gomori G (1953) Oxidative reactions of myeloid elements. J Histochem Cytochem 1:486–492
Graham GS (1916) The oxidizing ferment of the myelocyte series of cells and its demonstration by an alphanaphthol-pyronin method. J Med Res 35:231–242
Graham RS Jr, Karnovsky M (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney, ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302
Graham RC Jr, Lundholm V, Karnovsky MJ (1965) Cytochemical demonstration of peroxidase activity with 3-amino-9-ethylcarbazole. J Histochem Cytochem 13:150–152
Hanker JS, Yates PE, Metz CB, Rustioni A (1977) A new specific, sensitive and noncarcinogenic reagent for the demonstration of horseradish peroxidase. Histochem J 9:789–792
Hawley GG (1981) The condensed chemical dictionary. 10th edn. Van Nostrand Reinhold, New York
Hsu SM, Raine L, Fanger H (1981) The use of avidin-biotin peroxidase complex (ABC) in immunoperoxidase technique: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580
Jakobsen P, Andersen AP, Lyon H (1984) Preparation and characterization of methyl green tetrafluoroborate. Histochemistry 81:177–179
Lillie RD (1977) Conn's biological stains. Williams and Wilkins, Baltimore
Lillie RD, Burtner HJ (1953) Stable sudanophilia of human neutrophil leukocytes in relation to peroxidase and oxidase. J Histochem Cytochem 1:8–26
Loele W (1926) Uber oxidierende Substanzen in tierischen Zellen. Virchows Arch 261:484–502
Mauro A, Bertolotto A, Germano I, Giaccone G, Giordana MT, Migheli A, Schiffer D (1984) Collagenase in the immunohistochemical demonstration of laminin, fibronectin and factor VIII/RAg in nervous tissue after fixation. Histochemistry 80:157–163
Nakane PK (1968) Simultaneous localization of multiple tissue antigens using the peroxidase-labeled antibody method: a study on pituitary glands of the rat. J Histochem Cytochem 16:557–560
Pearse AGE (1972) Histochemistry. Theoretical and applied. Vol 2. 3rd edn. Churchill Livingstone, London
Pearse AGE (1980) Histochemistry. Theoretical and applied. Vol 1. 4th edn. Churchill Livingstone, London
Sax NI (1979) Dangerous properties of industrial materials. 5th edn. Van Nostrand Reinhold, New York
Sax NI (1981) Cancer causing chemicals. Van Nostrand Reinhold, New York
Seabra P (1955) Oxidase and lipase of the leukocyte. Ann NY Acad Sci 59, (art 5):1022–1051
Sofroniew MV, Schrell U (1982) Long term storage and regular repeated use of diluted antisera in glass staining jars for increased sensitivity, reproducibility and convenience of simple and two color light microscopic immunocytochemistry. J Histochem Cytochem 30:504–511
Sternberger LA, Hardy PH, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method for immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333
Van Norden S, Polak JM (1983) Immunocytochemistry today. Techniques and practice. In: Polak JM, Van Norden S (eds) Immunocytochemistry. Practical applications in pathology and biology. Wright PSG, Bristol, p 26
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Partially supported by a grant of the Italian National Research council, special Project “Oncology”, contract n. 84.00796.44 and by the Italian Association for Cancer Research (A.I.R.C.)
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Mauro, A., Germano, I., Giaccone, G. et al. 1-Naphthol basic dye (1-NBD). Histochemistry 83, 97–102 (1985). https://doi.org/10.1007/BF00495137
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DOI: https://doi.org/10.1007/BF00495137