Summary
A major drawback of all acrylic resins commonly used for post-embedding immunocytochemical studies of the central nervous system is the disruption of the ultrastructural morphology, due to the high lipid content of neural tissue. We have investigated the suitability of the acrylic resin LR Gold, which has been employed recently for immunogold labeling studies in several non-neural tissues. Optimal preservation of both antigenicity and ultrastructure of nervous tissue was obtained after en bloc staining with uranyl acetate, followed by total dehydration in acetone and curing at low temperature. Cell membranes and myelin sheaths, which are usually lost with other acrylic resins, were well maintained. The degree of antigenicity of LR Gold-embedded tissues was comparable to that of LR White-embedded one, but the morphologic detail was much better preserved. The use of LR Gold is particularly advantageous for studying neurodegenerative disorders such as Alzheimer disease.
Similar content being viewed by others
References
Bendayan M, Nanci A, Kan FWK (1987) Effect of tissue processing on colloidal gold cytochemistry. J Histochem Cytochem 35:983–996
Berryman MA, Rodewald RD (1990) An enhanced method for post-embedding immunocytochemical staining which preserves cell membranes. J Histochem Cytochem 38:159–170
Bettica A, Johnson AB (1990) Ultrastructural immunogold labeling of glial filaments in osmicated and unosmicated epoxy-embedded tissue. J Histochem Cytochem 38:103–109
Carlemalm E, Garavito RM, Villinger W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc 126:123–143
Castaño EN, Ghiso J, Prelli F, Gorevic PD, Migheli A, Frangione B (1986) In vitro formation of amyloid fibrils from two synthetic peptides of different lengths homologous to Alzheimer's disease β-protein. Biochem Biophys Res Commun 141:782–789
Erickson PA, Anderson DH, Fisher SK (1987) Use of uranyl acetate en bloc to improve tissue preservation and labeling for post-embedding immunoelectron microscopy. J Electron Microsc Tech 5:303–314
Goto M, Meyermann R, Wekerle H (1987) Ultrastructural immunocytochemistry of glia cells. Double labeling studies using LR White embedding and colloidal gold. Histochemistry 87:201–207
Gräber MB, Kreutzberg GW (1985) Immunogold staining (IGS) for electron microscopical demonstration of glial fibrillary acidic (GFA) protein in LR White embedded tissue. Histochemistry 83:497–500
Hayat A (1989a) Colloidal gold. Principles, methods, and applications. Academic Press, San Diego, USA
Hayat MA (1989b) Principles and techniques of electron microscopy. Biological applications, 3rd edn. Macmillan Press, Basingstoke, UK
Herken R, Fussek M, Thies M (1988) Light and electron microscopical postembedding lectin histochemistry for WGA-binding sites in the renal cortex of the mouse embedded in polyhydroxy aromatic resins LR-White and LR-Gold. Histochemistry 89:277–282
Lah JJ, Hayes DM, Burry RW (1990) A neutral pH silver development method for the visualization of 1-nanometer gold particles in pre-embedding electron microscopic immunocytochemistry. J Histochem Cytochem 38:503–508
Lamberts R, Goldsmith PC (1985) Preembedding colloidal gold immunostaining of hypothalamic neurons: light and electron microscopic localization of β-endorphin-immunoreactive perikarya. J Histochem Cytochem 33:499–507
Merighi A, Polak JM, Fumagalli G, Theodosis DT (1989) Ultrastructural localization of neuropeptides and GABA in rat dorsal horn: a comparison of different immunogold labeling techniques. J Histochem Cytochem 37:529–540
Migheli A, Attanasio A (1991) Two methods for flat embedding sections in LR White cut from paraffin blocks. Biotech Histochem 1:89–92
Migheli A, Mocellini C (1990) Ultrastructural immunocytochemistry in glial tumors. J Neurosurg Sci 34:219–222
Migheli A, Butler A, Brown K, Shelanski ML (1988) Light and electron microscope localization of the microtubule-associated tau protein in rat brain. J Neurosci 8:1846–1851
Migheli A, Mocellini C, Attanasio A (1991) Immunogold labeling of GFAP in osmicated, epoxy-embedded tissue. J Histochem Cytochem 39:879–880
Newman GR, Hobot JA (1987) Modern acrylics for post-embedding immunostaining techniques. J Histochem Cytochem 35:971–981
Pettitt JM, Humphris C (1991) Double lectin and immunolabelling for transmission electron microscopy: pre- and post-embedding application using the biotin-streptavidin system and colloidal gold-silver staining. Histochem J 23:29–37
Roth J (1989) Postembedding labeling on Lowicryl K4M tissue sections: detection and modification of cellular components. Method Cell Biol, vol. 31. Academic Press, San Diego, USA, pp 513–551
Somogyi P, Hodgson AJ (1985) Antisera to gamma-aminobutirric acid. III. Demonstration of GABA in Golgi-impregnated neurons and in conventional electron microscopic sections of cat striate cortex. J Histochem Cytochem 33:249–257
Stirling JW (1990) Immuno- and affinity probes for electron microscopy: a review of labeling and preparation techniques. J Histochem Cytochem 38:145–157
Valentino KL, Crumrine DA, Reichardt LF (1985) Lowicryl K4M embedding of brain tissue for immunogold electron microscopy. J Histochem Cytochem 33:969–973
Van den Pol AN (1984) Colloidal gold and biotin-avidin conjugates as ultrastructural markers for neuronal antigens. Q J Exp Psysiol 69:1–33
Yamaguchi H, Haga C, Hirai S, Nakazato Y, Kosaka K (1990) Distinctive, rapid, and easy labeling of diffuse plaques in the Alzheimer brains by a new methenamine silver stain. Acta Neuropathol 79:569–572
Yokota S, Oda T (1985) Immunocytochemical demonstration of serine: pyruvate aminotransferase in peroxisomes and mitochondria of rat kidney. Histochemistry 83:81–85
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Migheli, A., Attanasio, A. & Schiffer, D. LR Gold embedding of nervous tissue for immunoelectron microscopy studies. Histochemistry 97, 413–419 (1992). https://doi.org/10.1007/BF00270388
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00270388