Use of Immunogold with Silver Enhancement
- Constance Oliver
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Although gold particles are readily detectable by transmission electron microscopy, they can be difficult to visualize by bright-field light microscopy. If the particle size is large enough and the labeling dense enough, the gold particles will stain tissue red (1, 2). However, unless the gold is silver-enhanced to help visualize it, the sensitivity of the staining is fairly low. During silver-enhancement, the colloidal gold serves as a nucleation site for the deposition of metallic silver. The silver layer increases the size of the gold and imparts a black color to the stained tissue when viewed by bright-field microscopy (Fig. 1A). The silverenhanced gold particles can also be visualized using epipolarization, where they appear bright against a dark background (Fig. 1B). The silver enhancement method has its basis in 19th-century photographic techniques. The enhancing solutions are physical developers that contain both silver ions and a reducing agent, buffered to an acid pH. The developers most commonly used contain silver lactate as the source of silver ions (3). The silver lactate has a low dissociation coefficient that allows for more control of the reduction. Hydroquinone ( 1,4-dihydroxybenzene) is the only reducing agent that has been used in silver-enhancement techniques. A protective colloid, such as gum arabic, bovine serum albumin, dextran, polyethylene glycol (PEG), or polyvinylpyrrolidone (PVP), is frequently added in order to inhibit the autocatalytic reaction between the silver salt and the reducing agent.
- DeMey, J., Moeremans, M, Geuens, G., Nuydens, R., and DeBrander, M. (1981) High resolution light and electron microscopic localization of tubulin with the IGS (immuno-gold staining) method. Cell Biol. Int. Rep. 5, 889–899 CrossRef
- Geoghegan, W. D., Scillian, J. J., and Ackerman, G. A. (1978) The detection of human lymphocytes by both light and electron microscopy utilizing colloidal gold labeled anti-immunoglobulin. Immunol. Commun. 1, 1–12.
- Danscher, G. (1981) Histochemical demonstration of heavy metals. A revised version of the sulphide silver method suitable for both light and electron microscopy. Histochemistry 71, 1–16. CrossRef
- Roberts, W. J. (1935) A new procedure for the detection of gold in animal tissues: Physical development Proc. R. Acad. Sci. Amsterdam 38, 540–544.
- Danscher, G. (1981) Localization of gold in biological tissue. A photochemical method for light and electron microscopy Histochemistry 71, 81–88. CrossRef
- Holgate, C S., Jackson, P. I., Cowen, P. N., and Bird, C C (1983) Immunogoldsilver staining: new method of immunostaining with enhanced sensitivity. J. Histochem Cytochem. 31, 938–944
- Holgate, C S., Jackson, P., Lauder, I., Cowen, P., and Bird, C C (1983) Surface membrane staining of immunoglobulins in paraffin sections of non-Hodgkin’s lymphomas using immunogold-silver staining technique. J Clin Pathol. 36, 742–746 CrossRef
- Goode, D. and Maugel, T. K. (1987) Backscattered electron imaging of immunogold labeled and silver-enhanced microtubules in cultured mammalian cells. J. Electron Microsc. Tech. 5, 263–273. CrossRef
- deHarven, E. (1989) Backscattered electron imaging of the colloidal gold marker on cell surfaces, in Colloidal Gold, vol. 1 (Hayat, M. A., ed.), Academic, New York, pp. 229–249.
- Larsson, L-I (1988) Immunocytochemical detection systems, in Immunocytochemistry: Theory and Practice (Larsson, L.-I., ed), CRC, Boca Raton, FL, pp 77–146.
- Scopsi, L. (1989) Silver-enhanced colloidal gold method, in Colloidal Gold, vol. 1 (Hayat, M. A., ed.), Academic, New York, pp. 251–295.
- Hacker, G. W. (1989) Silver-enhanced colloidal gold for light microscopy, in Colloidal Gold, vol. 1 (Hayat, M A, ed), Academic, New York, pp. 297–321.
- Use of Immunogold with Silver Enhancement
- Book Title
- Immunocytochemical Methods and Protocols
- pp 211-216
- Print ISBN
- Online ISBN
- Series Title
- Methods in Molecular Bilogy
- Series Volume
- Series ISSN
- Humana Press
- Copyright Holder
- Humana Press
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