Summary
Elastin components have been identified in chick aorta by different immunoelectronmicroscopic procedures (peroxidase-antiperoxidase, immunoferritin and immunogold) using affinity purified antibodies to chick tropoelastin. The PAP method used in a preembedding procedure stained the outer portion of amorphous elastin and the microfibrils very intensively. The surface of the cells was also slightly stained. On the contrary immunogold labelling on Epon or Lowicryl embedded sections produced a strong decoration only of amorphous elastin, while microfibrils remained almost completely unlabelled. The result is not due to loss of antigenicity of microfibrils during embedding, since similar data were obtained with immunoferritin in a preembedding procedure. Experiments performed under different stringency conditions showed that the products of the peroxidase reaction diffuse and redistribute in the tissue, indicating that the positive staining of microfibrils and cell surface is an artifact. The value of different immunological reagents and procedures in studying the fine mapping of clastin components is discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Carlemalm E, Garavito RM, Villiger W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc (London) 126:123–143
Courtoy PJ, Picton DH, Farquhar MG (1983) Resolution and limitations of immunoperoxidase procedure in the localization of extracellular matrix antigens. J Histochem Cytochem 31:945–951
Daga-Gordini D, Bressan GM, Castellani I, Volpin D (1987) Fine mapping of tropoelastin-derived components in the aorta of developing chick embryo. Histochem J (in press)
Damiano VV, Tsang A, Christer P, Rosenbloom J, Weinbaum G (1979) Immunologic localization of elastin by electron microscopy. Am J Pathol 96:439–456
Daynes RA, Thomas M, Alvarez VL, Sandberg LB (1977) The antigenicity of soluble porcine elastins: I. Measurement of antibody by a radioimmunoassay. Connect Tissue Res 5:75–85
Fahrenbach WH, Sandberg LB, Cleary EG (1966) Ultrastructural studies on early elastogenesis. Anat Rec 155:573–576
Franc S, Garrone R, Bosch A, Franc JM (1984) A routine method for contrasting elastin at the ultrastructural level. J Histochem Cytochem 32:251–258
Fukuda Y, Ferrans VJ (1984) The electron microscopic immunohistochemistry of elastase-treated aorta and nuchal ligament of fetal and postnatal sheep. J Histochem Cytochem 32:747–756
Fukuda Y, Ferrans VJ, Crystal RG (1984) Development of elastic fibers of nuchal ligament, aorta and lung of fetal and postnatal sheep: an ultrastructural and electron microscopic study. Am J Anat 170:597–629
Geuze HJ, Slot JW, van der Ley P, Scheffer RCT (1981) Use of colloidal gold particles in double-labeling immunoelectron-microscopy of ultrathin frozen tissue sections. J Cell Biol 89:653–665
Horisberger M, Rosset J (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy. J Histochem Cytochem 25:295–305
Keeley FW (1981) Identification of elastin in developing aortic tissue by immunological methods. Connect Tissue Res 8:193–198
Lethias C, Hartmann DJ, Masmejean M, Ravazzola M, Sabbagh I, Ville G, Herbage D, Eloy R (1987) Ultrastructural immunolocalization of elastic fibers in rat blood vessels using the protein A-gold technique. J Histochem Cytochem 35:15–21
Miyata K, Takaya K (1981) Elastic fiber associated with collagenous fibrils surrounded by reticular cells in lymph nodes of the rat as revealed by electron microscopy after orcein staining. Cell Tissue Res 220:445–448
Novikoff AB (1980) DAB cytochemistry: artifacts problems in its current use. J Histochem Cytochem 28:1036–1041
Pasquali-Ronchetti I, Fornieri C, Castellani I, Bressan GM, Volpin D (1981) Alteration of the connective tissue components induced by β-aminopropionitrile. Exp Mol Pathol 35:42–56
Pasquali-Ronchetti I, Bressan GM, Fornieri C, Baccarani-Contri M, Castellani I, Volpin D (1984) Elastin fiber-associated glycosaminoglycans in β-aminopropionitrile-induced lathyrism. Exp Mol Pathol 40:235–245
Ross R, Bornstein P (1969) The elastic fiber. I. The separation and partial characterization of its macromolecular components. J Cell Biol 40:366–381
Roth J, Bendayan M, Carlemalm E, Villiger W, Garavito M (1981) The enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue. J Histochem Cytochem 29:663–671
Seligman AM, Shannon WA Jr, Hoshino Y, Plapinger RE (1973) Some important principles in 3, 3′-diaminobenzidine ultrastructural cytochemistry. J Histochem Cytochem 21:756–758
Sternberger LA (1979) Immunocytochemistry. 2nd edn. John Wiley and Sons, New York, pp 104–169
Yu SY, Sun CN, Still MF (1977) Ultrastructural changes of elastic tissue in hamster lung during elastase emphysema. Adv Exp Med Biol 79:39–56
Author information
Authors and Affiliations
Additional information
Supported by grant 86.00612.44 of the “Progetto Finalizzato Oncologia” of the Italian CNR
Rights and permissions
About this article
Cite this article
Daga-Gordini, D., Bressan, G.M., Castellani, I. et al. Detection of elastin by immunoelectronmicroscopy. Histochemistry 87, 573–578 (1987). https://doi.org/10.1007/BF00492473
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00492473